Cytoplasmic male sterile rudbeckia plants and methods for producing same

Abstract

To provide a cytoplasmic male sterile Rudbeckia plant.SOLUTION: A method for imparting cytoplasmic male sterility to a Rudbeckia plant is provided, including the step of introducing, to a Rudbeckia plant of interest, at least one polynucleotide having a specific sequence as a cytoplasmic male sterile gene.SELECTED DRAWING: Figure 6

Description

[Technical Field]

[0001] The present disclosure relates to cytoplasmically male sterile Rudbeckia plants and methods for producing same. [Background technology]

[0002] The genus Rudbeckia (hereinafter referred to as "Rudbeckia plants") is a plant with about 30 species that grow naturally in North America. The Rudbeckia plants are known to exist as annuals, biennials, and perennials (perennial plants). Among the Rudbeckia plants, the annual Rudbeckia sieboldii ( Rudbeckia hirta ) has a wide variety of flower colors, shapes, etc., and has been selectively bred for ornamental purposes, with many varieties being commercially available (Non-Patent Document 1).

[0003] Commercially available varieties of horticultural crops, including ornamental plants, are dominated by F1 varieties that utilize a phenomenon known as hybrid vigor. Hybrid vigor is a phenomenon in which the traits exhibited by the first generation hybrid (F1) are superior to the corresponding traits of either parent. The creation of F1 varieties requires the development of two parent lines of the F1 variety, followed by crossing and seed production between those lines. Furthermore, in order to prevent the contamination of self-fertilized seeds of the parent lines, methods such as artificial removal of stamens, use of self-incompatibility, and use of male sterility (MS) are used to produce F1 varieties.

[0004] Furthermore, in flowering plants, male sterility is not only used for the purpose of efficient seed production of F1 varieties, but can also provide ornamental benefits. Specifically, male sterile varieties do not release pollen from the anthers. Therefore, such male sterile varieties can prevent pollen from adhering to petals, etc., thereby preventing damage to the appearance of the plant. Furthermore, such male sterile varieties can prevent damage caused by pollen adhering to clothing. Furthermore, it has been reported that imparting male sterility to some flowering plants, such as petunias and pelargoniums, delays the aging of flowers that have opened, thereby extending the period during which they can be enjoyed (Non-Patent Documents 2 and 3).

[0005] Male sterility (MS) is classified into cytoplasmic male sterility (CMS) and karyotypic male sterility (GMS). In cytoplasmic male sterility, the male sterility gene is present in the cytoplasmic genome, i.e., the mitochondrial genome. On the other hand, in karyotypic male sterility, the male sterility gene is present in genomic DNA. [Prior art documents] [Non-patent literature]

[0006] [Non-Patent Document 1] Irene E. Palmer et al., “Crossability, Cytogenetics, and Reproductive Pathways in Rudbeckia Subgenus Rudbeckia”, HORTSCIENCE, 2009, Vol. 44, Issue 1 pages 44-48 [Non-patent document 2] Aran G. Smith et al., “Increased Flower Longevity in Petunia with Male Sterility”, HORTSCIENCE, 2004, Vol. 39, Issue 4, pages 822B-822 [Non-patent document 3] Begona Garcia-Sogo et al., “Production of engineered long-life and male sterile Pelargonium plants”, BMC Plant Biology, 2012, Vol. 12, Article number: 156 Summary of the Invention [Problem to be solved by the invention]

[0007] In Rudbeckia plants, no male sterility genes have been identified, and no cultivars exhibit male sterility. Therefore, an object of the present disclosure is to provide a Rudbeckia plant that exhibits cytoplasmic male sterility. [Means for solving the problem]

[0008] To achieve the above objectives, the Rudbeckia plants of the present disclosure are cytoplasmic male sterility.

[0009] A progeny line of the present disclosure is a progeny line of the Rudbeckia plant of the present disclosure, wherein the progeny line exhibits cytoplasmic male sterility.

[0010] The method for producing a cytoplasmically male sterile Rudbeckia plant of the present disclosure (hereinafter also referred to as the "first production method") comprises the following step (a): (a) crossing the Rudbeckia plant of the present disclosure or a progeny line of the present disclosure with another Rudbeckia plant.

[0011] The method of the present disclosure for imparting cytoplasmic male sterility to a Rudbeckia plant (hereinafter also referred to as the "imparting method") comprises the step of introducing into a target Rudbeckia plant at least one polynucleotide selected from the group consisting of the following (ca) to (ce) and (cf) as a cytoplasmic male sterility gene: (ca) any one of the following polynucleotides (ca1) to (ca7): (ca1) a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1; (ca2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which 1 to 67 bases are deleted, substituted, inserted and / or added in the base sequence of (ca1); (ca3) a polynucleotide that has a base sequence that is 90% or more identical to the base sequence of (ca1), and that causes cytoplasmic male sterility; (ca4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (ca1); (ca5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 2; (ca6) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence in which 1 to 22 amino acids are deleted, substituted, inserted, and / or added in the amino acid sequence of SEQ ID NO: 2; (ca7) a polynucleotide that expresses cytoplasmic male sterility and that consists of an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 2; (cb) any one of the following polynucleotides (cb1) to (cb7): (cb1) a polynucleotide consisting of the base sequence of SEQ ID NO: 3; (cb2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which 1 to 280 bases are deleted, substituted, inserted and / or added in the base sequence of (cb1); (cb3) a polynucleotide that expresses cytoplasmic male sterility and that consists of a base sequence having 90% or more identity to the base sequence of (cb1); (cb4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cb1); (cb5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 4; (cb6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 4 in which 1 to 93 amino acids have been deleted, substituted, inserted, and / or added; (cb7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 4; (cc) any one of the following polynucleotides (cc1) to (cc7): (cc1) a polynucleotide consisting of the base sequence of SEQ ID NO: 5; (cc2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which 1 to 126 bases are deleted, substituted, inserted and / or added in the base sequence of (cc1); (cc3) a polynucleotide that expresses cytoplasmic male sterility and that consists of a base sequence having 90% or more identity to the base sequence of (cc1); (cc4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to the polynucleotide having the nucleotide sequence of (cc1); (cc5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 6; (cc6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 6 in which 1 to 42 amino acids have been deleted, substituted, inserted, and / or added; (cc7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 6; (cd) any one of the following polynucleotides (cd1) to (cd7): (cd1) a polynucleotide consisting of the base sequence of SEQ ID NO: 7; (cd2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which 1 to 291 bases are deleted, substituted, inserted and / or added in the base sequence of (cd1); (cd3) a polynucleotide consisting of a base sequence having 90% or more identity to the base sequence of (cd1), which expresses cytoplasmic male sterility; (cd4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cd1); (cd5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 8; (cd6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 8 in which 1 to 97 amino acids have been deleted, substituted, inserted, and / or added; (cd7) a polynucleotide that expresses cytoplasmic male sterility and consists of an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 8; (ce) any one of the following polynucleotides (ce1) to (ce7): (ce1) a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 9; (ce2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which 1 to 114 bases are deleted, substituted, inserted and / or added in the base sequence of (ce1); (ce3) a polynucleotide that expresses cytoplasmic male sterility and that consists of a base sequence having 90% or more identity to the base sequence of (ce1); (ce4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the nucleotide sequence of (ce1); (ce5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 10; (ce6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 10 in which 1 to 38 amino acids have been deleted, substituted, inserted, and / or added; (ce7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 10; (cf) any one of the following polynucleotides (cf1) to (cf7): (cf1) a polynucleotide consisting of the base sequence of SEQ ID NO: 11; (cf2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which 1 to 46 bases are deleted, substituted, inserted and / or added in the base sequence of (cf1); (cf3) a polynucleotide that expresses cytoplasmic male sterility and that consists of a base sequence having 90% or more identity to the base sequence of (cf1); (cf4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to the polynucleotide having the nucleotide sequence of (cf1); (cf5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 12; (cf6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 12 in which 1 to 15 amino acids have been deleted, substituted, inserted, and / or added; (cf7) A polynucleotide consisting of an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 12, which expresses cytoplasmic male sterility.

[0012] The method for producing a cytoplasmic male sterile Rudbeckia plant of the present disclosure (hereinafter also referred to as the "second production method") includes a step of imparting cytoplasmic male sterility to a target Rudbeckia plant, The applying step is carried out by the applying method of the present disclosure.

[0013] The screening method for cytoplasmic male sterile Rudbeckia plants disclosed herein (hereinafter also referred to as the "screening method") includes a step (selection step) of selecting, from test Rudbeckia plants, test Rudbeckia plants that contain at least one polynucleotide selected from the group consisting of (ca) to (ce) and (cf) as a cytoplasmic male sterility gene, as cytoplasmic male sterile Rudbeckia plants.

[0014] The method for producing a cytoplasmic male sterile Rudbeckia plant of the present disclosure (hereinafter also referred to as the "third production method") includes a screening step of screening a test Rudbeckia plant containing a cytoplasmic male sterility gene from test Rudbeckia plants, The screening step is carried out by the screening method of the present disclosure.

[0015] The method for detecting cytoplasmic male sterility in Rudbeckia plants disclosed herein (hereinafter also referred to as the "detection method") includes a detection step of detecting at least one polynucleotide selected from the group consisting of (ca) to (ce) and (cf) as a cytoplasmic male sterility gene in a test Rudbeckia plant. [Effects of the Invention]

[0016] According to the present disclosure, a Rudbeckia plant exhibiting cytoplasmic male sterility can be provided. [Brief explanation of the drawings]

[0017] [Figure 1] FIG. 1 is a photograph showing the flowers of the Rudbeckia plant in Example 1. [Figure 2] FIG. 2 is a photograph of the Rudbeckia plant of the F1_MS1 line in Example 1. [Figure 3] FIG. 3 is a graph comparing the mitochondrial genomes of each lineage constructed by the two programs in Example 1. [Figure 4] FIG. 4 shows genes predicted on the mitochondrial genome of each strain constructed using Organella_PBA or HGAP4 in Example 1. [Figure 5] FIG. 5 is a schematic diagram comparing the base sequences of ATP6 of the RdMF strain and ORF3 of the RdCMS strain in Example 1. [Figure 6] FIG. 6 is a photograph showing the results of electrophoresis of the fertile lines (R. hiruta, RdMF line) in Example 1, the deposited line (RdCMS line), R. hiruta (120 lines) owned by Takii Seed Co., Ltd., and R. hiruta individuals growing wild within Takii Seed Co., Ltd. DETAILED DESCRIPTION OF THE INVENTION

[0018] <Definition> As used herein, "male sterility" or "male sterility" refers to a trait in which pollen formation does not occur or is insufficient due to the sterility of male organs. Specific examples of "male sterility" or "male sterility" include incomplete formation of stamens and loss of pollen due to underdevelopment.

[0019] As used herein, "cytoplasmic male sterility" or "cytoplasmic male sterility" refers to the trait of male sterility that is maternally inherited by genes derived from the cytoplasm or organelles present in the cytoplasm.

[0020] As used herein, "karyotypic male sterility" or "karyotypic male sterility" refers to the trait of male sterility inherited by genes of nuclear origin.

[0021] As used herein, "Rudbeckia plants" refers to plants belonging to the Asteraceae family ( Asteraceae ) of the genus Lamium ( Rudbeckia ) refers to plants classified as

[0022] As used herein, "cultivated Rudbeckia plants," "cultivated Rudbeckia cultivars," or "cultivated Rudbeckia" refer to agronomically superior Rudbeckia plants cultivated by humans, or their cultivars, breeding lines, or cultivars. "cultivated Rudbeckia plants," "cultivated Rudbeckia cultivars," or "cultivated Rudbeckia" may also be hybrids thereof, or hybrids with closely related species or wild species.

[0023] As used herein, the term "plant body" or "plant" refers to an individual plant that refers to the entire plant.

[0024] As used herein, "plant part" or "plant part" refers to a part of an individual plant.

[0025] As used herein, the term "polynucleotide" refers to a polymer of deoxyribonucleotides (DNA) and / or ribonucleotides (RNA). The polynucleotide may be a single-stranded polynucleotide or a double-stranded polynucleotide. The "polynucleotide" may also be referred to as, for example, a nucleic acid or a nucleic acid molecule.

[0026] As used herein, "polypeptide" refers to a polymer composed of unmodified amino acids (natural amino acids) and / or modified amino acids. The polypeptide is, for example, a peptide having a length of 10 amino acids or more. The "polypeptide" can also be referred to as, for example, a protein.

[0027] As used herein, "crossbreeding" refers to the crossing of two parental lines. Said crossbreeding can also be referred to as, for example, mating.

[0028] As used herein, an "expression vector" (vector) refers to in In vitro or in vivo In the context of the present invention, it refers to a recombinant plasmid or virus that contains the polynucleotide to be delivered to a host cell.

[0029] As used herein, "express" means that a desired trait is expressed in a subject or that a desired trait is induced in a subject.

[0030] The present disclosure will be described below using examples, but the present disclosure is not limited to the following examples and can be implemented with any modifications. Furthermore, the descriptions in this disclosure are mutually applicable unless otherwise specified. In this specification, when the expression "to" is used, it is used to mean the numerical or physical values ​​before and after it. In this specification, when multiple upper and / or lower limit values ​​are exemplified in a numerical range, any of the exemplified upper and lower limit values ​​can be combined as the upper and lower limit values ​​of the numerical range. In this specification, the expression "A and / or B" includes "A only," "B only," and "both A and B."

[0031] <Cytoplasmic male sterile Rudbeckia plant> In one aspect, the present disclosure provides a Rudbeckia plant that exhibits cytoplasmic male sterility. The Rudbeckia plant of the present disclosure exhibits cytoplasmic male sterility. The Rudbeckia plant of the present disclosure is characterized by exhibiting cytoplasmic male sterility, and other configurations and conditions are not particularly limited.

[0032] As a result of intensive research, the present inventors have discovered a novel Rudbeckia individual that exhibits cytoplasmic male sterility. As a result of further research, the present inventors have determined that in the cytoplasmic male sterile Rudbeckia individual, the cytoplasmic male sterility trait is a hereditary trait and is caused by a novel cytoplasmic male sterility gene. Therefore, according to the present disclosure, it is possible to provide a Rudbeckia plant that exhibits cytoplasmic male sterility.

[0033] In the Rudbeckia plants of the present disclosure, the male sterility is cytoplasmic male sterility (CMS). As described above, male sterility can be categorized as cytoplasmic male sterility or karyotypic male sterility (GMS). The type of male sterility is determined, for example, by the type of genome in which the male sterility gene is located (located). Specifically, if the male sterility gene is located in the cytoplasm, i.e., the mitochondrial genome, the Rudbeckia plant containing the male sterility gene exhibits cytoplasmic male sterility. On the other hand, if the male sterility gene is located in the nucleus, i.e., the nuclear genome, the Rudbeckia plant containing the male sterility gene exhibits karyotypic male sterility. Therefore, the type of male sterility can be evaluated, for example, by detecting the genome in which the male sterility gene is located. Specifically, if the male sterility gene is present in the mitochondrial genome of a test Rudbeckia plant, the test Rudbeckia plant can be evaluated as cytoplasmic male sterile, whereas if the male sterility gene is present in the nuclear genome of a test Rudbeckia plant, the test Rudbeckia plant can be evaluated as karyotypic male sterile.

[0034] The Rudbeckia plant may be, for example: Rudbeckia alpicola , Rudbeckia auriculata , Rudbeckia California , Rudbeckia Flava , Rudbeckia Fulgida , Rudbeckia glaucescens , Rudbeckia graminifolia , Rudbeckia Grandiflor , Rudbeckia heliopsidis , Rudbeckia hirta , Rudbeckia klamathensis , Rudbeckia laciniata , Rudbeckia missouriensis , Rudbeckia mohrii , Rudbeckia Mollis , Rudbeckia Montana , Rudbeckia newmannii , Rudbeckia nitida , Rudbeckia occidentalis , Rudbeckia scabrifolia , Rudbeckia speciosa , Rudbeckia subtomentosa , Rudbeckia texas , Rudbeckia triloba , Echinacea atrorubens , Echinacea pallida , Echinacea purpurea The Rudbeckia plant can also be referred to as, for example, a cultivated plant or cultivated variety of Rudbeckia plant.

[0035] The Rudbeckia plant may be a hybrid of a closely related species or a wild species. The closely related species may be, for example, a species of the genus Echinacea ( Echinacea ) plants, or Rudbeckia plants of a different species from the Rudbeckia plants to be hybridized.

[0036] Examples of the plant parts include plant cells, plant protoplasts, plant cell cultures or tissue cultures capable of regenerating plants, callus (plant callus), plant clumps, plant cells isolated from plants or plant parts, meristematic cells, pollen, flowers, petals, flower buds, corollas, leaves, petioles, leaf pith, cotyledons, ovaries, embryos, ovules, hypocotyls, egg cells, anthers, pistils, cuttings, scions, rootstocks, roots, root tips, seeds, fruits, stems, stalks, and seedlings. Examples of the plant parts include organs, tissues, cells, and vegetative propagules, and any of these may be used. Examples of the organs include petals, corollas, flowers, leaves, seeds, fruits, stems, and roots. Examples of the tissues are parts of the organs. The plant part may be, for example, the cytoplasm, mitochondria, or mitochondrial genome of an individual plant or its progeny; or a part of the plant such as a seed or callus.

[0037] The male sterility of the Rudbeckia plant is preferably a genetic trait. In this case, the male sterility of the Rudbeckia plant is caused, for example, by a male sterility gene. The male sterility gene may exist in the form of RNA (e.g., mRNA) or DNA (e.g., cDNA, genomic DNA, or mitochondrial DNA). The DNA may be double-stranded or single-stranded. The male sterility gene may also contain an additional sequence, such as a sequence of an untranslated region (UTR).

[0038] In the present disclosure, examples of the cell sterility gene include ORF3, ORF1, ORF2, ORF6, ORF7, and RPS7.

[0039] In the present disclosure, the ORF3 comprises the following polynucleotide (ca): (ca) any one of the following polynucleotides (ca1) to (ca7): (ca1) a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1; (ca2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (ca1); (ca3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (ca1), and that causes cytoplasmic male sterility; (ca4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (ca1); (ca5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 2; (ca6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 2 in which one or several amino acids have been deleted, substituted, inserted and / or added; (ca7) A polynucleotide consisting of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 2, which expresses cytoplasmic male sterility.

[0040] In (ca1), the nucleotide sequence of SEQ ID NO: 1 is a coding sequence for the amino acid sequence of SEQ ID NO: 2. The nucleotide sequence of SEQ ID NO: 1 can be obtained, for example, from seeds of the Rudbeckia plant deposited under accession number FERM BP-22428, which will be described later. The nucleotide sequence of SEQ ID NO: 1 below is a nucleotide sequence including a stop codon (TAA). As will be described later, the nucleotide sequence of SEQ ID NO: 1 is presumed to be the nucleotide sequence of a chimeric gene between the ATP6 gene of a fertile Rudbeckia plant and another gene. However, the present disclosure is in no way limited to this presumption.

[0041] Nucleotide sequence of the cytoplasmic male sterility gene (ORF3) (SEQ ID NO: 1) 5'--3'

[0042] In (ca2), "one or several" may be, for example, within a range in which the polynucleotide of (ca2) expresses the cytoplasmic male sterility. The "one or several" in (ca2) refers to, for example, 1 to 134, 1 to 100, 1 to 67, 1 to 33, 1 to 26, 1 to 20, 1 to 13, 1 to 6, 1 to 3, 1 or 2, or 1 in the base sequence of (ca1). In the present disclosure, a numerical range for the number of bases or amino acids, etc., discloses, for example, all positive integers within that range. For example, the description "1 to 5" means the disclosure of all of "1, 2, 3, 4, and 5" (the same applies hereinafter).

[0043] In the (ca3), the "identity" may be within a range in which the polynucleotide of the (ca3) expresses the cytoplasmic male sterility. The identity of the (ca3) to the base sequence of the (ca1) is, for example, 80% or more, 85% or more, 90% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more. The "identity" can be determined by aligning two base sequences or amino acid sequences (the same applies hereinafter). The alignment can be calculated using, for example, BLAST, FASTA, or the like with default parameters.

[0044] In (ca4), the "hybridizing polynucleotide" is, for example, a polynucleotide that is completely or partially complementary to the polynucleotide of (ca1). The "hybridizing polynucleotide" may be any polynucleotide as long as the polynucleotide of (ca4) expresses the cytoplasmic male sterility. The hybridization can be detected by, for example, various hybridization assays. The hybridization assay is not particularly limited, and may be, for example, a hybridization assay described in "Molecular Cloning: A Laboratory Manual 2nd Edition" edited by Sambrook et al. ndIt is also possible to employ the method described in "Cold Spring Harbor Laboratory Press (1989)" (Ed.)

[0045] In (ca4) above, "stringent conditions" may be, for example, low stringency conditions, moderate stringency conditions, or high stringency conditions. "Low stringency conditions" are, for example, 5x SSC, 5x Denhardt's solution, 0.5% SDS, 50% formamide, and 32°C. "Medium stringency conditions" are, for example, 5x SSC, 5x Denhardt's solution, 0.5% SDS, 50% formamide, and 42°C. "High stringency conditions" are, for example, 5x SSC, 5x Denhardt's solution, 0.5% SDS, 50% formamide, and 50°C. Those skilled in the art can set the degree of stringency by appropriately selecting conditions such as temperature, salt concentration, probe concentration and length, ionic strength, and time. The "stringent conditions" are, for example, those described in the aforementioned "Molecular Cloning: A Laboratory Manual 2nd Edition" edited by Sambrook et al. nd The conditions described in "Cold Spring Harbor Laboratory Press (1989)" can also be used.

[0046] The polynucleotide (ca5) may have, for example, a base sequence that allows the protein encoded by the polynucleotide (ca5) to express the cytoplasmic male sterility. The base sequence of the polynucleotide (ca5) can be designed, for example, by substituting corresponding codons based on the amino acid sequence of SEQ ID NO: 2. The amino acid sequence of SEQ ID NO: 2 can be obtained, for example, from seeds of the Rudbeckia plant deposited under accession number FERM BP-22428, which will be described later.

[0047] Amino acid sequence encoded by the cytoplasmic male sterility gene (ORF3) (SEQ ID NO: 2) MLLTLSLVLLLIHFVTKKGGGNLVPNAWQSLVELIYDFVLNLVNEQIGGLSGNVKQKFFPCILVTFTFLLFCNLQGMIPYSFTVTSHFLITLGLSFSIFIGITIVGFQRNG LHFLSFLLPAGVPLPLAPFLVLLELISYCFRALSLGIRLFANMMAGHSLVKILSGFAWTMLCMNDLLYFIGDLGPLFIVLALTGLELGVAILQAYVFTILICIYLNDAINLH

[0048] In the (ca6) amino acid sequence, the "one or several" may be within a range in which the polynucleotide of (ca6) expresses the cytoplasmic male sterility. The "one or several" in the (ca6) amino acid sequence may be, for example, 1 to 44, 1 to 33, 1 to 22, 1 to 11, 1 to 8, 1 to 6, 1 to 4, 1 to 3, 1 or 2, or 1 in the amino acid sequence of SEQ ID NO: 2.

[0049] The "identity" of the amino acid sequence of (ca7) may be within a range in which the polynucleotide of (ca7) expresses the cytoplasmic male sterility. The identity of (ca7) to the amino acid sequence of SEQ ID NO: 2 is, for example, 80% or more, 85% or more, 90% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more.

[0050] The phrase "to express cytoplasmic male sterility" means, for example, that in a plant individual containing a target polynucleotide, the plant individual exhibits cytoplasmic male sterility (the same applies hereinafter).

[0051] In the present disclosure, the ORF1 comprises the polynucleotide (cb) below. (cb) any one of the following polynucleotides (cb1) to (cb7): (cb1) a polynucleotide consisting of the base sequence of SEQ ID NO: 3; (cb2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cb1); (cb3) a polynucleotide that expresses cytoplasmic male sterility and that consists of a base sequence having 80% or more identity to the base sequence of (cb1); (cb4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cb1); (cb5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 4; (cb6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 4 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cb7) A polynucleotide consisting of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 4, which expresses cytoplasmic male sterility.

[0052] In (cb1), the nucleotide sequence of SEQ ID NO: 3 is a coding sequence for the amino acid sequence of SEQ ID NO: 4. The nucleotide sequence of SEQ ID NO: 3 can be obtained, for example, from seeds of the Rudbeckia plant deposited under accession number FERM BP-22428, which will be described later. The nucleotide sequence of SEQ ID NO: 3 below is a nucleotide sequence that does not contain a stop codon (TAA). As will be described later, the nucleotide sequence of SEQ ID NO: 3 is presumed to be a DNA-dependent RNA polymerase of the Rudbeckia plant. However, the present disclosure is in no way limited to this presumption.

[0053] Nucleotide sequence of the cytoplasmic male sterility gene (ORF1) (SEQ ID NO: 3)

[0054] In (cb2), the "one or several" may be, for example, within a range in which the polynucleotide of (cb2) expresses the cytoplasmic male sterility. The "one or several" in (cb2) may be, for example, 1 to 561, 1 to 421, 1 to 280, 1 to 140, 1 to 112, 1 to 84, 1 to 56, 1 to 28, 1 to 26, 1 to 20, 1 to 13, 1 to 6, 1 to 3, 1 or 2, or 1 in the base sequence of (cb1).

[0055] In the (cb3), the "identity" may be within a range in which the polynucleotide of the (cb3) expresses the cytoplasmic male sterility. The identity of the (cb3) to the base sequence of the (cb1) is, for example, 80% or more, 85% or more, 90% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more.

[0056] In (cb4), the "hybridizing polynucleotide" is, for example, a polynucleotide that is completely or partially complementary to the polynucleotide in (cb1). The "hybridizing polynucleotide" may be any polynucleotide that allows the polynucleotide in (cb4) to express the cytoplasmic male sterility. The explanation of hybridization in (ca4) can be applied to the hybridization.

[0057] The polynucleotide (cb5) may have, for example, a base sequence that allows the protein encoded by the polynucleotide (cb5) to express the cytoplasmic male sterility. The base sequence of the polynucleotide (cb5) can be designed, for example, by substituting corresponding codons based on the amino acid sequence of SEQ ID NO: 4. The amino acid sequence of SEQ ID NO: 4 can be obtained, for example, from seeds of the Rudbeckia plant deposited under accession number FERM BP-22428, which will be described later.

[0058] Amino acid sequence encoded by the cytoplasmic male sterility gene (ORF1) (SEQ ID NO: 4) MEQTVREFLLSTVSLDDNKKKGIVVDFWSEFYQKNVYTEHQSNQTNRSLYKEKVCEILHEFKRTHITPYSREELIALQAEIESTTIIFDEASIHASSGHIMKYMIDPKKESAKDFVYERLTSRKEQSIISDMGQYTLEAIIVYVISKLYSSETEMIRVSTLIDQLDKQVKAQSVLVNKHKKPLVMKEVGQNVALGEHFPIGAALVEFISDRGLMTIQHIDDRRSSVPIQKKKGKYYMPKLLYAFYNFDVSLLPVKLNIPMVFPPIEWSNAREDNLEPKTLYDLRGGYISGMSEVSRYHLLSSWNYNNYYIDIESGHESLCVVMNKLQKVPFRIASFMLTFIKENYNDFVKAGLLMPKALCIADMKKLRDSLRDFYMNNEKGIKTIYSYDEVLTILYKDVQRARFERMTLMLSEALEGFKLYLPAFLDFRGRIYRSGILHLHERDFVRSLLIFDNLYDYKNQDEIIHINKCLSILGRAIPYYWKSFSSYTDSEEWGRKLIEIKDNICNSELIKFSVSAKKPFQFLASVCALKIEDETTRNHYLNYLPITQDASSSAYQIMSFLLLDTQIAKQTNLISEHKGEIHDIYNHMKELLLEYIESCSGEYDLSENLKELVIKSIDRKLVKALFMPIIYGKTVISTATDLRTTLSKFVTKGESMILAKICFSFWKETYRGMDCLITLIKSIGWMVSARGSHVRYQNQIYTTVQKYMKMDAVKIWIYDRENKKKRQVTLRVSTEKNDKRKSEIATFVNFIHQFDAFIAMSVIYEMPSYASHIYTVHDNFITIPSSCDDLPYLYKKAFSCNNPLAIINRYIYTNVMEHLGMIEGLTPVESDHFRRLRKKDGYDTLIIHENILLKYLICNMPINLKRKNETQMWEGKINTIITSYKEYVRTICMCEDPGKDYDINKLEKCRDEYTENDQNFIRRMMRGSTNYSIHH

[0059] In the (cb6) amino acid sequence, the "one or several" may be within a range in which the polynucleotide of (cb6) expresses the cytoplasmic male sterility. The "one or several" in the (cb6) amino acid sequence may be, for example, 1 to 187, 1 to 140, 1 to 93, 1 to 46, 1 to 35, 1 to 28, 1 to 18, 1 to 9, 1 to 8, 1 to 6, 1 to 4, 1 to 3, 1 or 2, or 1 in the amino acid sequence of SEQ ID NO: 4.

[0060] The "identity" of the amino acid sequence of (cb7) may be within a range in which the polynucleotide of (cb7) expresses the cytoplasmic male sterility. The identity of (cb7) to the amino acid sequence of SEQ ID NO: 4 is, for example, 80% or more, 85% or more, 90% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more.

[0061] In the present disclosure, the ORF2 comprises the following polynucleotide (cc): (cc) any one of the following polynucleotides (cc1) to (cc7): (cc1) a polynucleotide consisting of the base sequence of SEQ ID NO: 5; (cc2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cc1); (cc3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (cc1), and that expresses cytoplasmic male sterility; (cc4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to the polynucleotide having the nucleotide sequence of (cc1); (cc5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 6; (cc6) A polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 6 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cc7) A polynucleotide consisting of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 6, which expresses cytoplasmic male sterility.

[0062] In (cc1), the nucleotide sequence of SEQ ID NO: 5 is a coding sequence for the amino acid sequence of SEQ ID NO: 6. The nucleotide sequence of SEQ ID NO: 5 can be obtained, for example, from seeds of the Rudbeckia plant deposited under accession number FERM BP-22428, which will be described later. The nucleotide sequence of SEQ ID NO: 5 below is a nucleotide sequence that does not contain a stop codon (TAA). As will be described later, the nucleotide sequence of SEQ ID NO: 5 is presumed to be a DNA polymerase of the Rudbeckia plant. However, the present disclosure is in no way limited to this presumption.

[0063] Nucleotide sequence of cytoplasmic male sterility gene (ORF2) (SEQ ID NO: 5)

[0064] In (cc2), the "one or several" may be, for example, within a range in which the polynucleotide of (cc2) expresses the cytoplasmic male sterility. The "one or several" in (cc2) may be, for example, 1 to 253, 1 to 190, 1 to 126, 1 to 63, 1 to 50, 1 to 38, 1 to 25, 1 to 12, 1 to 6, 1 to 3, 1 or 2, or 1 in the base sequence of (cc1).

[0065] In the (cc3), the "identity" may be within a range in which the polynucleotide of the (cc3) expresses the cytoplasmic male sterility. The identity of the (cc3) to the base sequence of the (cc1) is, for example, 80% or more, 85% or more, 90% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more.

[0066] In (cc4), the "hybridizing polynucleotide" is, for example, a polynucleotide that is completely or partially complementary to the polynucleotide in (cc1). The "hybridizing polynucleotide" may be any polynucleotide that expresses cytoplasmic male sterility in (cc4). The explanation of hybridization in (ca4) can be applied to the hybridization.

[0067] The polynucleotide (cc5) may have, for example, a base sequence that allows the protein encoded by the polynucleotide (cc5) to express the cytoplasmic male sterility. The base sequence of the polynucleotide (cc5) can be designed, for example, by substituting corresponding codons based on the amino acid sequence of SEQ ID NO: 6. The amino acid sequence of SEQ ID NO: 6 can be obtained, for example, from seeds of the Rudbeckia plant deposited under accession number FERM BP-22428, which will be described later.

[0068] Amino acid sequence encoded by the cytoplasmic male sterility gene (ORF2) (SEQ ID NO: 6) MLGFIEAYVVCPKTIKKPFLPYREKEGTLLFPTGEFVGVYFSEELKYAREIGYTVIPISGYLFEKKESPFREFVSDLFESRLEAKKSGNDALSYVYKILMNSLYG RFGINPMGTITEICDSKRHKLLIRKTELISTDELTDSKYIVTYRSNTETDYWDPPKNSAVQIAAAITAYARIYMYPYISREDSYYTDTDSVVLGQPLSDELISSSI LGKFKLEDKIVEGYFLAPKSYYYRNDKGEDILKYKGLAKTEITPEWFRSQYANPDRTLEVEVEANFRIDWSSLNIFKKDKNVKVGLNQNPKRIKVYEGKYWVDTMP VDVKDLSRLDNISRKLVTWLKADVTHLKNENLSLNEKLSEKEREITEKKRMDERDNEMKEEPTEVTNPTIDIDEIPKIDIDEIPKMKPKKKAKTDKKTTTTKKKKPP

[0069] In the (cc6) amino acid sequence, the "one or several" may be within a range in which the polynucleotide of (cc6) expresses the cytoplasmic male sterility. The "one or several" in (cc6) may be, for example, 1 to 86, 1 to 63, 1 to 42, 1 to 21, 1 to 16, 1 to 12, 1 to 8, 1 to 6, 1 to 4, 1 to 3, 1 or 2, or 1 in the amino acid sequence of SEQ ID NO: 6.

[0070] In the (cc7), the "identity" of the amino acid sequence may be within a range in which the polynucleotide of the (cc7) expresses the cytoplasmic male sterility. The identity of the (cc7) to the amino acid sequence of SEQ ID NO: 6 is, for example, 80% or more, 85% or more, 90% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more.

[0071] In the present disclosure, the ORF6 comprises the following polynucleotide (cd): (cd) any one of the following polynucleotides (cd1) to (cd7): (cd1) a polynucleotide consisting of the base sequence of SEQ ID NO: 7; (cd2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cd1); (cd3) a polynucleotide consisting of a base sequence having 80% or more identity to the base sequence of (cd1), which expresses cytoplasmic male sterility; (cd4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cd1); (cd5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 8; (cd6) A polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 8 in which one or several amino acids have been deleted, substituted, inserted and / or added; (cd7) A polynucleotide consisting of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 8, which expresses cytoplasmic male sterility.

[0072] In (cd1), the nucleotide sequence of SEQ ID NO: 7 is a coding sequence for the amino acid sequence of SEQ ID NO: 8. The nucleotide sequence of SEQ ID NO: 7 can be obtained, for example, from seeds of the Rudbeckia plant deposited under accession number FERM BP-22428, which will be described later. The nucleotide sequence of SEQ ID NO: 7 below is a nucleotide sequence that does not contain a stop codon (TAA). As will be described later, the nucleotide sequence of SEQ ID NO: 7 is presumed to be a DNA polymerase of the Rudbeckia plant. However, the present disclosure is in no way limited to this presumption.

[0073] Nucleotide sequence of cytoplasmic male sterility gene (ORF6) (SEQ ID NO: 7)

[0074] In (cd2), the "one or several" may be within a range in which the polynucleotide of (cd2) expresses the cytoplasmic male sterility. The "one or several" in (cd2) may be, for example, 1 to 583, 1 to 437, 1 to 291, 1 to 145, 1 to 116, 1 to 87, 1 to 58, 1 to 50, 1 to 38, 1 to 29, 1 to 25, 1 to 12, 1 to 6, 1 to 3, 1 or 2, or 1 in the base sequence of (cd1).

[0075] In the (cd3), the "identity" may be within a range in which the polynucleotide of the (cd3) expresses the cytoplasmic male sterility. The identity of the (cd3) to the base sequence of the (cd1) is, for example, 80% or more, 85% or more, 90% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more.

[0076] In the (cd4) polynucleotide, the "hybridizing polynucleotide" is, for example, a polynucleotide that is completely or partially complementary to the (cd1) polynucleotide. The "hybridizing polynucleotide" may be any polynucleotide that allows the (cd4) polynucleotide to express the cytoplasmic male sterility. The explanation of the hybridization in the (ca4) polynucleotide can be applied to the hybridization.

[0077] The polynucleotide (cd5) may have, for example, a base sequence that allows the protein encoded by the polynucleotide (cd5) to express the cytoplasmic male sterility. The base sequence of the polynucleotide (cd5) can be designed, for example, by substituting corresponding codons based on the amino acid sequence of SEQ ID NO: 8. The amino acid sequence of SEQ ID NO: 8 can be obtained, for example, from seeds of the Rudbeckia plant deposited under accession number FERM BP-22428, which will be described later.

[0078] Amino acid sequence encoded by the cytoplasmic male sterility gene (ORF6) (SEQ ID NO: 8) MNKMNMKMKRIMPMCSRTLHTSVRPELEFVFNRYQSVVEQTRVQDPHPGLIIATLNFKKPGLGVDDIELISIATMDLLNQYVYPSISGYGKFTISLRMIQSIEEEITYTIGCAIPLTSNDGTLLPKNEIYARIKEAYQKNAELYNGCSLVQLIIRAYLDKVERLDRPELTVSDRYEELLSIQSDKLSEIEAISARKIQHSKRQYREYITRIKRVSSGKKAFIVSDLETIQIDYKHRPYAAGLMLVREGKDIKDSLIYTYFSEDYSVYIKSFEKRSQKVLFDLVRKIIALSKIERSAMTVYFHNFSRFDGIILLKHLACHHDYKLKQLFRNNRLYELKVYSGRKLLFKMRDSLNLLPGKLDNLAKSLCPSLGGKGSLNYDDVRADNLVSKKDQLISYMKQDILLLGGIMKKAQEIYYDLYQLDIVSKITLSSLALSIYRMRYYDEENWPIYIPNMNQDHFIRKAYYGGHTDVYKPYGENLYYYDVNSLYPFVMKNFQMPGGQPVWHGNLLDKDLDSLYGFIEAYVVCPKTINKPFLPYRNKNNTLIFPTGEFAGVYYSEELKFARDLGYTVLPLSGYLYERMESPFKEFVNTQSSKRIEAKKEGNDALSYVYKILMNSLYGRFGINPKSTTSEICDHDRYVKMLKDDSFLQGSLLDKNKYIVIYHVNTGSNPESWNPPKNGAVQLAAAITACARIYMYPLISREDCYYTDTDSVVLGQPLSDELISASELGMLKLEARILKGYFLAPKSYAYIQYDENKEIVIKHKGAAKNLVTMEWFQSQYDDPSRTQLVSVTSNFKINWNELEIHKQETLYRLGISQDSKRLPVYCEKKWIDTEPIDIRDLSNHSPQMLDRILAYLRDEVNRHQTNSEILRKELSKKDSEMISIISDKDRVISEMKSRIESLQEMRKITNPTDKTEKKTHTAKKTKTEKKTHTAKKTKTDKKKTTNQTLKKHQNQRKQRPPRNHHTTKKPP

[0079] In the (cd6), the "one or several" in relation to the amino acid sequence may be within a range in which the (cd6) polynucleotide expresses the cytoplasmic male sterility. The "one or several" in the (cd6) may be, for example, 1 to 194, 1 to 145, 1 to 97, 1 to 48, 1 to 38, 1 to 29, 1 to 18, 1 to 16, 1 to 12, 1 to 9, 1 to 8, 1 to 6, 1 to 4, 1 to 3, 1 or 2, or 1 in the amino acid sequence of SEQ ID NO: 8.

[0080] The "identity" of the amino acid sequence of (cd7) may be within a range in which the polynucleotide of (cd7) expresses the cytoplasmic male sterility. The identity of (cd7) to the amino acid sequence of SEQ ID NO: 8 is, for example, 80% or more, 85% or more, 90% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more.

[0081] In the present disclosure, the ORF7 comprises the polynucleotide (ce) below. (ce) any one of the following polynucleotides (ce1) to (ce7): (ce1) a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 9; (ce2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (ce1); (ce3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (ce1), and that causes cytoplasmic male sterility; (ce4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the nucleotide sequence of (ce1); (ce5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 10; (ce6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 10 in which one or several amino acids have been deleted, substituted, inserted and / or added; (ce7) A polynucleotide consisting of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 10, which expresses cytoplasmic male sterility.

[0082] In (ce1), the nucleotide sequence of SEQ ID NO: 9 is a coding sequence for the amino acid sequence of SEQ ID NO: 10. The nucleotide sequence of SEQ ID NO: 9 can be obtained, for example, from seeds of the Rudbeckia plant deposited under accession number FERM BP-22428, which will be described later. The nucleotide sequence of SEQ ID NO: 9 below is a nucleotide sequence that does not contain a stop codon (TAA).

[0083] Nucleotide sequence of cytoplasmic male sterility gene (ORF7) (SEQ ID NO: 9)

[0084] In (ce2), the "one or several" may be within a range in which the polynucleotide of (ce2) expresses the cytoplasmic male sterility. The "one or several" in (ce2) may be, for example, 1 to 229, 1 to 171, 1 to 114, 1 to 57, 1 to 45, 1 to 34, 1 to 22, 1 to 11, 1 to 6, 1 to 3, 1 or 2, or 1 in the base sequence of (ce1).

[0085] In the (ce3), the "identity" may be within a range in which the polynucleotide of the (ce3) expresses the cytoplasmic male sterility. The identity of the (ce3) to the base sequence of the (ce1) is, for example, 80% or more, 85% or more, 90% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more.

[0086] In (ce4), the "hybridizing polynucleotide" is, for example, a polynucleotide that is completely or partially complementary to the polynucleotide in (ce1). The "hybridizing polynucleotide" may be any polynucleotide that allows the polynucleotide in (ce4) to express the cytoplasmic male sterility. The explanation of hybridization in (ca4) can be applied to the hybridization.

[0087] The polynucleotide (ce5) may have, for example, a base sequence that allows the protein encoded by the polynucleotide (ce5) to express the cytoplasmic male sterility. The base sequence of the polynucleotide (ce5) can be designed, for example, by substituting corresponding codons based on the amino acid sequence of SEQ ID NO: 10. The amino acid sequence of SEQ ID NO: 10 can be obtained, for example, from seeds of the Rudbeckia plant deposited under accession number FERM BP-22428, which will be described later.

[0088] Amino acid sequence encoded by the cytoplasmic male sterility gene (ORF7) (SEQ ID NO: 10) MTNPVQRDQSKFLNNTVVMNDQKKKDVVVEFWKSFYGILFVSFTNFVLILLTCILVEPETVQMIARFIGGSSAIYFLFLARARFSKLFNLFSFLVTTCYLFVLNHLNAPDIGLVPICVCGCFIFSYYFTEKHTGWHFDLCFMIILTCRLIVSPDIWAISLVFELLVLFCLLDHDMDQDRMRLCCLYFILLL FLCCNYLYGSSIHIDTLVVVLVLAAAGAGGIQFMSLSKTERGETLGLQLFLINNVILGFLLRKDGETLPMITIFFIVSILCFCIGLYLIGRIKEQSFYSVLSELKSSLRGLFIPVLLTLNHFFLEDQYYLWITSLVTRIIVIQLLQLILKKEDEDPDQDKAIKSDDVRRCEINGCRSCYNYYYARSPPDCY

[0089] In the (ce6) amino acid sequence, the "one or several" may be within a range in which the polynucleotide of (ce6) expresses the cytoplasmic male sterility. The "one or several" in the (ce6) amino acid sequence may be, for example, 1 to 76, 1 to 57, 1 to 38, 1 to 19, 1 to 15, 1 to 12, 1 to 9, 1 to 8, 1 to 6, 1 to 4, 1 to 3, 1 or 2, or 1 in the amino acid sequence of SEQ ID NO: 10.

[0090] The "identity" of the amino acid sequence of (ce7) may be within a range in which the polynucleotide of (ce7) expresses the cytoplasmic male sterility, for example, and the identity of (ce7) to the amino acid sequence of SEQ ID NO: 10 is, for example, 80% or more, 85% or more, 90% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more.

[0091] In the present disclosure, the RPS7 comprises the polynucleotide (cf) below. (cf) any one of the following polynucleotides (cf1) to (cf7): (cf1) a polynucleotide consisting of the base sequence of SEQ ID NO: 11; (cf2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cf1); (cf3) a polynucleotide that expresses cytoplasmic male sterility and that consists of a base sequence having 80% or more identity to the base sequence of (cf1); (cf4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to the polynucleotide having the nucleotide sequence of (cf1); (cf5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 12; (cf6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 12 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cf7) A polynucleotide consisting of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 12, which polynucleotide causes expression of cytoplasmic male sterility.

[0092] In (cf1), the nucleotide sequence of SEQ ID NO: 11 is a coding sequence for the amino acid sequence of SEQ ID NO: 12. The nucleotide sequence of SEQ ID NO: 11 can be obtained, for example, from seeds of the Rudbeckia plant deposited under accession number FERM BP-22428, which will be described later. The nucleotide sequence of SEQ ID NO: 11 below is a nucleotide sequence including a stop codon (TAA).

[0093] Nucleotide sequence of the cytoplasmic male sterility gene (RPS7) (SEQ ID NO: 11) 5'-ATGTCACGTCGAGGTACTGCAGAAGAAAAACTGCAAAATCCGATCCAATTTATCGTAATCGATTAGTTAACATGTTGGTTAACCGTATTCTGAAAACACGGAAAAAAATCATTGGCTTATCAAATTATCTATCGAGCCGTGAAAAAGATTCAACAAAAGACAGAAACAAATCCACTATCTGTTTACGTCAAGCAATACATGGAGTAACTCCGGGTATAGCAGTAAAAGCAAGA CGGTGTAGGTGGATCGACTCATCAAGTTCCCATTGAAATAGGATCCACACAAGGAAAAGCACTTGCCATTCGTTGGTTATTAGCGGCATCCCGAAAACGTCCGGGTCGAAATATGGCTTTCAAATTAAGTTCCGAATTAGTGGATGCTGCCAAAGGGAGTGGCGATGCCATACGCAAAAGGGAAGAGACTCATAGAATGGCAGAGGCAAATAGAGCTTTTGCACATTTTCGTTAA-3'

[0094] In (cf2), the "one or several" may be, for example, within a range in which the polynucleotide of (cf2) expresses the cytoplasmic male sterility. The "one or several" in (cf2) may be, for example, 1 to 93, 1 to 70, 1 to 46, 1 to 23, 1 to 18, 1 to 14, 1 to 9, 1 to 6, 1 to 3, 1 or 2, or 1 in the base sequence of (cf1).

[0095] In the (cf3), the "identity" may be within a range in which the polynucleotide of (cf3) expresses the cytoplasmic male sterility. The identity of (cf3) to the base sequence of (cf1) is, for example, 80% or more, 85% or more, 90% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more.

[0096] In (cf4), the "hybridizing polynucleotide" is, for example, a polynucleotide that is completely or partially complementary to the polynucleotide in (cf1). The "hybridizing polynucleotide" may be any polynucleotide that expresses cytoplasmic male sterility in (cf4). The explanation of hybridization in (ca4) can be applied to the hybridization.

[0097] The polynucleotide (cf5) may have, for example, a base sequence that allows the protein encoded by the polynucleotide (cf5) to express the cytoplasmic male sterility. The base sequence of the polynucleotide (cf5) can be designed, for example, by substituting corresponding codons based on the amino acid sequence of SEQ ID NO: 12. The amino acid sequence of SEQ ID NO: 12 can be obtained, for example, from seeds of the Rudbeckia plant deposited under accession number FERM BP-22428, which will be described later.

[0098] Amino acid sequence encoded by the cytoplasmic male sterility gene (RPS7) (SEQ ID NO: 12) MSRRGTAEEKTAKSDPIYRNRLVNMLVNRILKHGKKSLAYQIIYRAVKKIQQKTETNPLSVLRQAIHGVTPGIAVKARRVGGSTHQVPIEIGSTQGKALAIRWLLAASRKRPGRNMAFKLSSELVDAAKGSGDAIRKREETHRMAEANRAFAHFR

[0099] In (cf6), the "one or several" in the amino acid sequence may be within a range in which the polynucleotide of (cf6) expresses the cytoplasmic male sterility. For example, the "one or several" in (cf6) may be, for example, 1 to 31, 1 to 23, 1 to 15, 1 to 7, 1 to 6, 1 to 4, 1 to 3, 1 or 2, or 1 in the amino acid sequence of SEQ ID NO: 12.

[0100] In the (cf7) polynucleotide, the "identity" of the amino acid sequence may be within a range in which the polynucleotide of the (cf7) expresses the cytoplasmic male sterility. The identity of the (cf7) polynucleotide to the amino acid sequence of SEQ ID NO: 12 is, for example, 80% or more, 85% or more, 90% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more.

[0101] In the present disclosure, the Rudbeckia plant may contain any one, multiple, or all of the cytoplasmic male sterility genes. When the Rudbeckia plant contains one cytoplasmic male sterility gene, the cytoplasmic male sterility gene preferably includes ORF3. When the Rudbeckia plant contains multiple cytoplasmic male sterility genes, the combinations of the multiple cytoplasmic male sterility genes may be, for example, the following combinations: Polynucleotides (ca) and (cb) Polynucleotides (ca) and (cc) Polynucleotides (ca) and (cd) Polynucleotides (ca) and (ce) Polynucleotides (ca) and (cf) Polynucleotides (cb) and (cc) Polynucleotides (cb) and (cd) Polynucleotides (cb) and (ce) Polynucleotides (cb) and (cf) Polynucleotides (cc) and (cd) Polynucleotides (cc) and (ce) Polynucleotides (cc) and (cf) (cd) and (ce) polynucleotides Polynucleotides (cd) and (cf) Polynucleotides (ce) and (cf) Polynucleotides (ca), (cb), and (cc) Polynucleotides (ca), (cb), and (cd) (ca), (cb), and (ce) polynucleotides Polynucleotides (ca), (cb), and (cf) (ca), (cc), and (cd) polynucleotides (ca), (cc), and (ce) polynucleotides Polynucleotides (ca), (cc), and (cf) (ca), (cd), and (ce) polynucleotides Polynucleotides (ca), (cd), and (cf) Polynucleotides (ca), (ce), and (cf) (cb), (cc), and (cd) polynucleotides (cb), (cc), and (ce) polynucleotides Polynucleotides (cb), (cc), and (cf) (cb), (cd), and (ce) polynucleotides Polynucleotides (cb), (cd), and (cf) (cb), (ce), and (cf) polynucleotides (cc), (cd), and (ce) polynucleotides (cc), (cd), and (cf) polynucleotides (cc), (ce), and (cf) polynucleotides (cd), (ce), and (cf) polynucleotides Polynucleotides (ca), (cb), (cc), and (cd) (ca), (cb), (cc), and (ce) polynucleotides Polynucleotides (ca), (cb), (cc), and (cf) (ca), (cb), (cd), and (ce) polynucleotides Polynucleotides (ca), (cb), (cd), and (cf) Polynucleotides (ca), (cb), (ce), and (cf) (ca), (cc), (cd), and (ce) polynucleotides (ca), (cc), (cd), and (cf) polynucleotides (ca), (cc), (ce), and (cf) polynucleotides (ca), (cd), (ce), and (cf) polynucleotides (cb), (cc), (cd), and (ce) polynucleotides (cb), (cc), (cd), and (cf) polynucleotides (cb), (cc), (ce), and (cf) polynucleotides (cb), (cd), (ce), and (cf) polynucleotides (cc), (cd), (ce), and (cf) polynucleotides (ca), (cb), (cc), (cd), and (ce) polynucleotides Polynucleotides (ca), (cb), (cc), (cd), and (cf) Polynucleotides (ca), (cb), (cc), (ce), and (cf) Polynucleotides (ca), (cb), (cd), (ce), and (cf) (ca), (cc), (cd), (ce), and (cf) polynucleotides (cb), (cc), (cd), (ce), and (cf) polynucleotides (ca), (cb), (cc), (cd), (ce), and (cf) polynucleotides

[0102] When the Rudbeckia plant of the present disclosure contains the cytoplasmic male sterility gene, the cytoplasmic male sterility gene is present in the cytoplasm or mitochondrial genome (mitochondrial DNA). In this case, the cytoplasmic male sterility gene may be an endogenous gene or an exogenous gene. The endogenous gene is, for example, a gene generated by a mutation or the like in the Rudbeckia plant. The exogenous gene is, for example, a gene introduced by genome editing technology or the like.

[0103] In the Rudbeckia plant of the present disclosure, the ORF3 is presumed to be a chimeric gene of the ATP6 gene and another gene, as described below. Therefore, when the Rudbeckia plant of the present disclosure contains the ORF3, it is preferable that the Rudbeckia plant of the present disclosure have the cytoplasmic male sterility gene instead of or as the ATP6 gene.

[0104] An example of a Rudbeckia plant exhibiting cytoplasmic male sterility is the Rudbeckia plant (deposited line) deposited under accession number FERM BP-22428 or its progeny. The deposit information is shown below. Hereinafter, the deposited line will also be referred to as Rudbeckia cultivar Takii 22. Type of deposit: International deposit Name of depository institution: National Institute of Technology and Evaluation, Patent Organism Depositary Center Address: Room 120, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, 292-0818, Japan Accession number: Accession number FERM BP-22428 Identification mark: Takii 22 Received: September 14, 2021

[0105] The deposited line may be a plant that is substantially the same as the deposited line to the extent that it has cytoplasmic male sterility. Examples of the substantially identical plant include mutants and genetically modified plants of the deposited line.

[0106] The Rudbeckia plants of the present disclosure can be obtained, for example, by directly or indirectly assessing and selecting cytoplasmic male sterility from a test Rudbeckia plant.

[0107] The direct assessment of cytoplasmic male sterility can be carried out, for example, based on the ability to form male gametophytes. Specifically, if the test Rudbeckia plant does not form stamens (male stamens), pollen, and / or anthers, preferably stamens and / or anthers, the test Rudbeckia plant can be assessed as male sterile. On the other hand, if the test Rudbeckia plant forms stamens, pollen, and / or anthers, preferably stamens and / or anthers, the test Rudbeckia plant can be assessed as not being male sterile, i.e., as male fertile.

[0108] The direct evaluation also evaluates the type of male sterility in the male-sterile test Rudbeckia plant, i.e., whether it is CMS or GMS. In this case, evaluation can be performed by backcrossing multiple times using the test Rudbeckia plant and its progeny line as seed parents and a male-fertile Rudbeckia plant as the pollen parent (revert parent). The pollen parent may or may not contain a fertility restorer, but it is preferable that it does not contain the fertility restorer, as this allows for more accurate evaluation of whether the plant is CMS or GMS. Examples of the fertility restorer include the PPR (Pentatricopeptide repeat) gene. If all individuals of the progeny line obtained by backcrossing are male sterile, the test Rudbeckia plant can be evaluated as CMS. On the other hand, if the progeny lines obtained by the backcrossing are male sterile and male fertile, the test Rudbeckia plant can be evaluated as GMS.

[0109] The indirect evaluation of cytoplasmic male sterility can be carried out, for example, based on the presence or absence of the cytoplasmic male sterility gene. Specifically, if a test Rudbeckia plant has the cytoplasmic male sterility gene, the test Rudbeckia plant can be evaluated as cytoplasmic male sterile. On the other hand, if a test Rudbeckia plant does not have the cytoplasmic male sterility gene, the test Rudbeckia plant can be evaluated as not male sterile, i.e., male fertile. A method for detecting the cytoplasmic male sterility gene will be described below.

[0110] The indirect assessment further includes assessing the type of male sterility in the test Rudbeckia plant, i.e., whether it is CMS or GMS. In this case, assessment can be performed by detecting the location of the cytoplasmic male sterility gene in the test Rudbeckia plant. If the cytoplasmic male sterility gene is present in the cytoplasm, mitochondria, or mitochondrial genome in the test Rudbeckia plant, the male sterility can be assessed as CMS. On the other hand, if the male sterility gene is present in the nucleus or nuclear genome in the test Rudbeckia plant, the male sterility can be assessed as GMS.

[0111] The indirect assessment of cytoplasmic male sterility may be carried out based on the presence or absence of the cytoplasmic male sterility gene in the cytoplasm, mitochondria, or mitochondrial genome. In this case, if the test Rudbeckia plant has the cytoplasmic male sterility gene in the cytoplasm, mitochondria, or mitochondrial genome, the test Rudbeckia plant can be assessed as cytoplasmic male sterile. On the other hand, if the test Rudbeckia plant does not have the cytoplasmic male sterility gene in the cytoplasm, mitochondria, or mitochondrial genome, the test Rudbeckia plant can be assessed as not being cytoplasmic male sterile, i.e., as being karyotypically cytoplasmic male sterile or male fertile. Methods for detecting the cytoplasmic male sterility gene will be described below.

[0112] In the present disclosure, the male-sterile Rudbeckia plant can be obtained by selecting a test Rudbeckia plant that has been evaluated as male sterile. In the present disclosure, a test Rudbeckia plant with CMS may be selected as the cytoplasmic male-sterile Rudbeckia plant.

[0113] The Rudbeckia plant of the present disclosure may be produced by introducing the cytoplasmic male sterility gene into a target Rudbeckia plant. In this case, the Rudbeckia plant of the present disclosure can be obtained by producing a transformant of the present disclosure using the vector of the present disclosure described below. In producing the transformant, the cytoplasmic male sterility gene can be introduced into the cytoplasm, mitochondria, or mitochondrial genome to produce the CMS Rudbeckia plant. In introducing the cytoplasmic male sterility gene, for example, the cytoplasmic male sterility gene may be introduced in addition to the ATP6 gene, or the cytoplasmic male sterility gene may be introduced instead of the ATP6 gene or by modifying the ATP6 gene.

[0114] The Rudbeckia plant of the present disclosure may be a progeny line of a Rudbeckia plant exhibiting cytoplasmic male sterility. The progeny line may be an individual plant of the progeny line, or a portion of an individual plant of the progeny line, a seed of the progeny line, a callus of the progeny line, the cytoplasm of the progeny line, mitochondria of the progeny line, or a mitochondrial genome of the progeny line. The progeny line may also be, for example, a Rudbeckia plant into which the cytoplasmic male sterility gene has been introgressed.

[0115] The progeny line may be a plant obtained by crossing a Rudbeckia plant of the present disclosure with another Rudbeckia plant, or a Rudbeckia plant of the present disclosure with a wild-caught Rudbeckia plant. The progeny line may be obtained, obtainable, or derived directly or indirectly by cross-pollinating a Rudbeckia plant of the present disclosure or its progeny line, or may be derived from a parent line obtained from a Rudbeckia plant of the present disclosure using traditional breeding methods such as cross-pollination. Examples of the progeny line include first-generation hybrid F1 (hybrid first-generation line, F1 hybrid), backcross progeny, etc. When obtaining the progeny line, a Rudbeckia plant of the present disclosure can be used as a female parent, and progeny lines can be obtained by cross-pollination.

[0116] The cross may be "cross-pollination" or "self-pollination." Cross-pollination refers to fertilization by the union of two gametes from different plants. Self-pollination refers to the transfer of pollen from anthers to the stigma of the same plant. Self-pollination can also be referred to as selfing, for example. The cross may include backcrossing, which is a traditional breeding method.

[0117] Backcrossing is a traditional breeding technique in which breeders repeatedly backcross progeny lines of a hybrid with one of the parent lines to introduce a trait into a plant or variety. A plant containing the trait to be introduced can be referred to as, for example, a donor plant. A plant into which the trait is introduced can be referred to as, for example, a recurrent parent. Backcrossing can be performed by crossing a donor plant with the recurrent parent, thereby obtaining a first-generation hybrid F1 (first-generation hybrid line, F1 hybrid). The progeny line possessing the trait is then crossed with the recurrent parent. The trait of the donor plant can then be introduced into the recurrent parent through several generations of backcrossing and / or selfing.

[0118] Rudbeckia plants of the present disclosure may possess other desirable traits in addition to the trait of cytoplasmic male sterility.

[0119] Rudbeckia plants of the present disclosure exclude, for example, plants obtained by essentially biological processes or plants obtained exclusively by essentially biological processes.

[0120] For the method of producing a cytoplasmic male sterile Rudbeckia plant of the present disclosure, the explanations of the imparting method, second production method, screening method, and third production method described below can be used.

[0121] The Rudbeckia plants of the present disclosure exhibit cytoplasmic male sterility. Therefore, the Rudbeckia plants of the present disclosure do not require emasculation of seed parents when obtaining first-generation hybrid lines, and highly pure seeds can be supplied. Therefore, the Rudbeckia plants of the present disclosure can reduce the effort required for breeding and harvesting. Furthermore, the Rudbeckia plants of the present disclosure can be easily maintained and propagated by crossing them with a maintainer line that has substantially the same traits other than cytoplasmic male sterility. Furthermore, the Rudbeckia plants of the present disclosure can suppress damage to external traits or clothing caused by pollen, and can enjoy a longer viewing period compared to Rudbeckia plants that have substantially the same traits other than cytoplasmic male sterility.

[0122] <First production method> In another aspect, the present disclosure provides a method for producing a cytoplasmic male sterile Rudbeckia plant. The method for producing a cytoplasmic male sterile Rudbeckia plant of the present disclosure, as described above, comprises the steps of: (a) growing a cytoplasmic male sterile Rudbeckia plant; (a) Crossing a cytoplasmically male sterile Rudbeckia plant of the present disclosure or its progeny line with another Rudbeckia plant.

[0123] The first production method of the present disclosure is characterized in that the Rudbeckia plant of the present disclosure is used in step (a), and other steps and conditions are not particularly limited. According to the first production method of the present disclosure, a Rudbeckia plant exhibiting cytoplasmic male sterility can be produced.

[0124] In step (a), the plant used as the first parent may be the cytoplasmically male sterile Rudbeckia plant of the present disclosure or a progeny thereof. The Rudbeckia plant of the present disclosure exhibits cytoplasmic male sterility. Therefore, in step (a), the Rudbeckia plant of the present disclosure or a progeny thereof is used as the seed parent (female parent). As described above, the cytoplasmically male sterile Rudbeckia plant of the present disclosure can also be obtained, for example, by the imparting method, second production method, screening method, and third production method of the present disclosure described below. Therefore, the first production method of the present disclosure may, for example, involve carrying out one or more of the imparting method, second production method, screening method, and third production method of the present disclosure prior to step (a). In this case, the explanation of each method can be referred to in the explanation of each method described below.

[0125] As a specific example, the first production method of the present disclosure may include the following step (x) or step (y): (x) A step of selecting the Rudbeckia plant of the present disclosure or its progeny line from the test Rudbeckia plant (selection step) (y) A step of producing the Rudbeckia plant of the present disclosure or its progeny line from the target Rudbeckia plant (production step)

[0126] In the step (x), the selection of the cytoplasmic male sterile Rudbeckia plant of the present disclosure or its progeny line can be carried out, for example, by directly or indirectly evaluating the cytoplasmic male sterility of the test Rudbeckia plant and selecting the cytoplasmic male sterile Rudbeckia plant of the present disclosure or its progeny line. The above-mentioned explanation of the method for evaluating cytoplasmic male sterility can be used for the direct evaluation.

[0127] When the indirect evaluation is carried out, the selection of a cytoplasmic male sterile Rudbeckia plant or its progeny line according to the present disclosure can be said to be the selection of a Rudbeckia plant having the cytoplasmic male sterility gene. In this case, the step (x) can be carried out, for example, by the following steps (x1) and (x2). (x1) a detection step of detecting the presence or absence of a cytoplasmic male sterility gene in the test Rudbeckia plant; (x2) A selection step of selecting the test Rudbeckia plant as a cytoplasmic male sterile Rudbeckia plant when the presence of the cytoplasmic male sterility gene, i.e., the cytoplasmic male sterility gene, is detected.

[0128] When step (x) includes steps (x1) and (x2), step (x) can be carried out, for example, using the base sequence of the cytoplasmic male sterility gene as an indicator. In step (x1), the cytoplasmic male sterility gene may be detected, for example, based on the presence or absence of a base sequence encoding the cytoplasmic male sterility gene. Furthermore, in step (x1), the cytoplasmic male sterility gene may be detected using a detection reagent for the cytoplasmic male sterility gene, such as a primer set and / or probe, that can identify the cytoplasmic male sterility gene.

[0129] When detecting the presence or absence of a nucleotide sequence encoding the cytoplasmic male sterility gene, the step (x1) involves, for example, decoding the nucleotide sequence of the mitochondrial genome (mitochondrial DNA) of the test Rudbeckia plant. Then, in the step (x1), the obtained nucleotide sequence is compared with the nucleotide sequence of the polynucleotide (ca), and detection can be performed based on whether the nucleotide sequences match. The nucleotide sequence can be decoded, for example, using a sample containing the mitochondrial genome (mitochondrial DNA), a sequencing reagent, and a sequencer. The comparison of the nucleotide sequences can be performed, for example, using nucleotide sequence analysis software (e.g., the aforementioned BLAST). The region of the obtained nucleotide sequence to be compared with the nucleotide sequence may be an intron region of the mitochondrial genome (mitochondrial DNA), an exon region of the mitochondrial genome (mitochondrial DNA), or both. Thus, in step (x1), a Rudbeckia plant having a base sequence that matches the base sequence of the cytoplasmic male sterility gene can be specified (identified or detected) as a test Rudbeckia plant containing the cytoplasmic male sterility gene. Then, in step (x2), for example, a test Rudbeckia plant containing the cytoplasmic male sterility gene is selected as a Rudbeckia plant of the present disclosure or its progeny, i.e., a Rudbeckia plant exhibiting cytoplasmic male sterility.

[0130] When a detection reagent for the cytoplasmic male sterility gene is used for detection, the cytoplasmic male sterility gene is detected in step (x1) using, for example, a primer set, a probe, or a combination thereof capable of detecting the cytoplasmic male sterility gene. Specifically, when the primer set is used, the cytoplasmic male sterility gene can be detected in step (x1) by, for example, performing PCR using a sample containing the mitochondrial genome (mitochondrial DNA) and the primer set, and determining whether an amplified fragment indicating the presence of the cytoplasmic male sterility gene is obtained. When the probe is used, the cytoplasmic male sterility gene can be detected in step (x1) by, for example, using a sample containing the mitochondrial genome (mitochondrial DNA) and the probe, and determining whether a probe-derived signal indicating the presence of the cytoplasmic male sterility gene is obtained. When the primer set and the probe are used, in the step (x1), for example, PCR is performed using a sample containing the mitochondrial genome (mitochondrial DNA) and the primer set to amplify an amplified fragment indicating the presence of the cytoplasmic male sterility gene. Next, in the step (x1), the cytoplasmic male sterility gene can be detected by using the amplified fragment and the probe to determine whether a probe-derived signal indicating the presence of the cytoplasmic male sterility gene is obtained. Then, in the step (x2), for example, a test Rudbeckia plant containing the cytoplasmic male sterility gene is selected as a Rudbeckia plant of the present disclosure or its progeny, i.e., a Rudbeckia plant exhibiting cytoplasmic male sterility.

[0131] In step (x1), it is sufficient to detect one or more of ORF3, ORF1, ORF2, ORF6, ORF7, and RPS7 as the cytoplasmic male sterility gene, or to detect a plurality of or all of them. In step (x1), ORF3 may be detected as the cytoplasmic male sterility gene, or ORF3, ORF1, ORF2, ORF6, ORF7, and RPS7 may be detected. When a plurality of cytoplasmic male sterility genes are detected in step (x1), test Rudbeckia plants in which one or more, a plurality, or all of the cytoplasmic male sterility genes are detected are selected in step (x2) as Rudbeckia plants or their progeny lines of the present disclosure, i.e., Rudbeckia plants exhibiting cytoplasmic male sterility.

[0132] The ORF3 is presumed to be a chimeric gene of the ATP6 gene and another gene. Therefore, when detecting ORF3 in step (x1), the primer set and / or probe is designed, for example, to be capable of distinguishing between the ORF3 and the ATP6 gene. The primer set may be, for example, a combination of a primer set capable of distinguishing between the ORF3 and the ATP6 gene. The nucleotide sequence of the ORF (open reading frame) of the ATP6 gene of the Rudbeckia plant can be, for example, the nucleotide sequence shown below (SEQ ID NO: 13, including a stop codon (TAA)). The ATP6 gene may be a functional equivalent as long as its functionality is maintained. In the nucleotide sequence shown below in SEQ ID NO: 13, the nucleotide sequence from 763 to 1512 is the same as the nucleotide sequence from 340 to 1089 in the nucleotide sequence shown in SEQ ID NO: 1. Therefore, in the present disclosure, primers and / or probes capable of amplifying different nucleotide sequences in the nucleotide sequences shown in SEQ ID NO: 1 and SEQ ID NO: 13 are designed. Then, in step (x1), the designed primers and / or probes can be used to detect whether the test Rudbeckia plant contains the ORF3 and / or ATP6 gene. The primer set and probe can be designed, for example, using known design methods. An example of a primer set capable of identifying ORF3 and a primer set capable of identifying the ATP6 gene is a combination of a primer set for ORF3 (a cytoplasmic male sterility gene) and a primer set for ATP6, as shown below. Thus, in step (x1), a test Rudbeckia plant in which ORF3 has been identified by the primer set and / or probe can be specified (identified or detected) as a test Rudbeckia plant containing the cytoplasmic male sterility gene. Then, in the step (x2), for example, a test Rudbeckia plant containing the ORF3 (cytoplasmic male sterility gene) is selected as a Rudbeckia plant of the present disclosure or its progeny line, i.e., a Rudbeckia plant exhibiting cytoplasmic male sterility.

[0133] Nucleotide sequence of the ORF of the ATP6 gene (SEQ ID NO: 13)

[0134] Primer set for ORF3 Forward primer (SEQ ID NO: 14) 5'- ACGTAACACGTATCTATGGTGCAT-3' Reverse primer (SEQ ID NO: 15) 5'-GAGAGTTAGCAGCATAAACAAAGA-3' Primer set for ATP6 Forward primer (SEQ ID NO: 16) 5'- GAGGGACTTTATTCAGTCTTATCG-3' Reverse primer (SEQ ID NO: 17) 5'- ATCTTCATAGGAATCAATGGGAGA-3'

[0135] When detecting ORF1, ORF2, ORF6, ORF7, and / or RPS7, primer sets and / or probes for each gene can be designed based on the base sequence of the cytoplasmic male sterility gene described above using conventional methods or software in the technical field of this application (e.g., Primer-BLAST, etc.).

[0136] In the step (x1), the location of the cytoplasmic male sterility gene may be detected. Examples of the location include the nucleus, nuclear genome, cytoplasm, mitochondria, and mitochondrial genome, and preferably the cytoplasm, mitochondria, or mitochondrial genome. In this case, in the step (x2), if the test Rudbeckia plant contains the cytoplasmic male sterility gene and the cytoplasmic male sterility gene is present in the cytoplasm, mitochondria, or mitochondrial genome, the test Rudbeckia plant can be selected as a Rudbeckia plant exhibiting cytoplasmic male sterility.

[0137] In step (x1), the sample for detecting the cytoplasmic male sterility gene may be, for example, a sample containing the nucleus, nuclear genome, cytoplasm, mitochondria, and / or mitochondrial genome. Each sample can be prepared from the test Rudbeckia plant or a part thereof by a conventional method in the technical field of the present application. The sample is preferably a sample containing the cytoplasm, mitochondria, and / or mitochondrial genome.

[0138] The step (y) can also be referred to as, for example, a step of introducing the cytoplasmic male sterility gene into the target Rudbeckia plant (an introduction step). The introduction step can be carried out using the same explanations as for the cytoplasmic male sterility gene, expression vector, transformant, and method of imparting cytoplasmic male sterility described below in the present disclosure.

[0139] Next, in the step (a), the Rudbeckia plant used as the other parent is not particularly limited, and may be, for example, a male-fertile Rudbeckia plant.

[0140] In the step (a), the method for crossbreeding the cytoplasmically male sterile Rudbeckia plant with the other Rudbeckia plant is not particularly limited, and any known method can be used.

[0141] In step (b), the target for selecting a cytoplasmically male sterile Rudbeckia plant may be, for example, the Rudbeckia plant obtained in step (a), or may be a progeny line obtained from that Rudbeckia plant. Specifically, the target may be, for example, an F1 Rudbeckia plant obtained by crossing in step (a), or a progeny line thereof. The progeny line may be, for example, a backcross progeny line of the F1 Rudbeckia plant obtained by crossing in step (a), or a Rudbeckia plant obtained by crossing a progeny line of the F1 Rudbeckia plant or the like with another Rudbeckia plant.

[0142] In the step (b), the selection of cytoplasmic male sterile Rudbeckia plants can be carried out, for example, by directly or indirectly assessing cytoplasmic male sterility. The above-described method for assessing cytoplasmic male sterility can be used for the direct assessment.

[0143] In step (b), the selection of the cytoplasmic male sterile Rudbeckia plant can be carried out, for example, by directly or indirectly evaluating the cytoplasmic male sterility of the obtained F1 Rudbeckia plant or its progeny line, and selecting the cytoplasmic male sterile Rudbeckia plant. When the selection is carried out by the indirect evaluation, the selection of the cytoplasmic male sterile Rudbeckia plant can be said to be selection of a Rudbeckia plant having the cytoplasmic male sterility gene. In this case, step (b) can be carried out, for example, by steps (b1) and (b2) described below. (b1) a detection step of detecting the presence or absence of a cytoplasmic male sterility gene in the obtained F1 Rudbeckia plant or its progeny line; (b2) A selection step in which, if the presence of the cytoplasmic male sterility gene, i.e., the cytoplasmic male sterility gene, is detected, the obtained F1 Rudbeckia plant or its progeny line is selected as a cytoplasmic male sterile Rudbeckia plant.

[0144] The selection of cytoplasmic male sterile Rudbeckia plants in step (b) can be carried out, for example, in the same manner as described in step (x), and step (b1) can be carried out in the same manner as step (x1), and step (b2) can be carried out in the same manner as step (x2).

[0145] In step (b1), the sample for detecting the cytoplasmic male sterility gene may be, for example, a sample containing the nucleus, nuclear genome, cytoplasm, mitochondria, and / or mitochondrial genome. Each sample can be prepared from the test Rudbeckia plant or a part thereof by a conventional method in the technical field of the present application. The sample is preferably a sample containing the cytoplasm, mitochondria, and / or mitochondrial genome.

[0146] In the first production method of the present disclosure, it is preferable to further cultivate the cytoplasmically male sterile Rudbeckia plant selected in step (b). The cultivation conditions and cultivation method for the Rudbeckia plant can be appropriately determined, for example, depending on the growth stage of the Rudbeckia plant and the variety of the Rudbeckia plant. During the cultivation, the Rudbeckia plant may be grown to any growth stage.

[0147] Thus, in the step (b), the Rudbeckia plant or its progeny line in which the cytoplasmic male sterility has been confirmed can be selected as a cytoplasmic male sterile Rudbeckia plant.

[0148] The first production method of the present disclosure may further include a seed collection step of collecting seeds from the progeny line obtained by crossbreeding.

[0149] <Cytoplasmic male sterility gene> In another aspect, the present disclosure provides a gene capable of conferring cytoplasmic male sterility. The cytoplasmic male sterility gene of the present disclosure comprises at least one polynucleotide selected from the group consisting of (ca) to (ce) and (cf) below.

[0150] (ca) any one of the following polynucleotides (ca1) to (ca7): (ca1) a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1; (ca2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (ca1); (ca3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (ca1), and that causes cytoplasmic male sterility; (ca4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (ca1); (ca5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 2; (ca6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 2 in which one or several amino acids have been deleted, substituted, inserted and / or added; (ca7) a polynucleotide that expresses cytoplasmic male sterility and that consists of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 2; (cb) any one of the following polynucleotides (cb1) to (cb7): (cb1) a polynucleotide consisting of the base sequence of SEQ ID NO: 3; (cb2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cb1); (cb3) a polynucleotide that expresses cytoplasmic male sterility and that consists of a base sequence having 80% or more identity to the base sequence of (cb1); (cb4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cb1); (cb5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 4; (cb6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 4 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cb7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 4; (cc) any one of the following polynucleotides (cc1) to (cc7): (cc1) a polynucleotide consisting of the base sequence of SEQ ID NO: 5; (cc2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cc1); (cc3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (cc1), and that expresses cytoplasmic male sterility; (cc4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to the polynucleotide having the nucleotide sequence of (cc1); (cc5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 6; (cc6) A polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 6 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cc7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 6; (cd) any one of the following polynucleotides (cd1) to (cd7): (cd1) a polynucleotide consisting of the base sequence of SEQ ID NO: 7; (cd2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cd1); (cd3) a polynucleotide consisting of a base sequence having 80% or more identity to the base sequence of (cd1), which expresses cytoplasmic male sterility; (cd4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cd1); (cd5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 8; (cd6) A polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 8 in which one or several amino acids have been deleted, substituted, inserted and / or added; (cd7) a polynucleotide that expresses cytoplasmic male sterility and consists of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 8; (ce) any one of the following polynucleotides (ce1) to (ce7): (ce1) a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 9; (ce2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (ce1); (ce3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (ce1), and that causes cytoplasmic male sterility; (ce4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the nucleotide sequence of (ce1); (ce5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 10; (ce6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 10 in which one or several amino acids have been deleted, substituted, inserted and / or added; (ce7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 10; (cf) any one of the following polynucleotides (cf1) to (cf7): (cf1) a polynucleotide consisting of the base sequence of SEQ ID NO: 11; (cf2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cf1); (cf3) a polynucleotide that expresses cytoplasmic male sterility and that consists of a base sequence having 80% or more identity to the base sequence of (cf1); (cf4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to the polynucleotide having the nucleotide sequence of (cf1); (cf5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 12; (cf6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 12 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cf7) A polynucleotide consisting of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 12, which polynucleotide causes expression of cytoplasmic male sterility.

[0151] The explanation of the polynucleotides (ca) to (cf) can be made in accordance with the explanation of the Rudbeckia plants of the present disclosure.

[0152] The cytoplasmic male sterility gene of the present disclosure may include one or more, a plurality, or all of the polynucleotides (ca) to (cf). When the cytoplasmic male sterility gene of the present disclosure includes a plurality of polynucleotides, the combination of the polynucleotides can be described in the same manner as in the combination of a plurality of cytoplasmic male sterility genes in the Rudbeckia plant of the present disclosure.

[0153] The cytoplasmic male sterility gene of the present disclosure can be suitably used for synthesizing (producing or breeding) the cytoplasmic male sterile Rudbeckia plant of the present disclosure by genetic engineering techniques. When the cytoplasmic male sterility gene of the present disclosure is composed of DNA, the cytoplasmic male sterility gene of the present disclosure can also be referred to as, for example, recombinant DNA.

[0154] The polynucleotide can be synthesized by, for example, genetic engineering techniques or organic synthesis techniques, and can also be called synthetic DNA such as cDNA or synthetic RNA.

[0155] <Cytoplasmic male sterility protein> In another aspect, the present disclosure provides a protein that is predicted to induce cytoplasmic male sterility. The cytoplasmic male sterility protein of the present disclosure comprises at least one polypeptide selected from the group consisting of (CA) to (CE) and (CF) below.

[0156] (CA) any one of the following polypeptides (CA1) to (CA3): (CA1) a polypeptide consisting of the amino acid sequence of SEQ ID NO: 2; (CA2) A polypeptide that expresses cytoplasmic male sterility, consisting of an amino acid sequence in which one or several amino acids are deleted, substituted, inserted and / or added in the amino acid sequence of SEQ ID NO: 2; (CA3) A polypeptide that expresses cytoplasmic male sterility, consisting of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 2; (CB) any one of the following polypeptides (CB1) to (CB3): (CB1) a polypeptide consisting of the amino acid sequence of SEQ ID NO: 4; (CB2) A polypeptide that expresses cytoplasmic male sterility, consisting of the amino acid sequence of SEQ ID NO: 4 in which one or several amino acids have been deleted, substituted, inserted and / or added; (CB3) A polypeptide that expresses cytoplasmic male sterility, consisting of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 4; (CC) any one of the following polypeptides (CC1) to (CC3): (CC1) a polypeptide consisting of the amino acid sequence of SEQ ID NO: 6; (CC2) A polypeptide that expresses cytoplasmic male sterility, consisting of the amino acid sequence of SEQ ID NO: 6 in which one or several amino acids have been deleted, substituted, inserted and / or added; (CC3) A polypeptide that expresses cytoplasmic male sterility, consisting of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 6; (CD) any one of the following polypeptides (CD1) to (CD3): (CD1) a polypeptide consisting of the amino acid sequence of SEQ ID NO: 8; (CD2) A polypeptide that expresses cytoplasmic male sterility, consisting of the amino acid sequence of SEQ ID NO: 8 in which one or several amino acids have been deleted, substituted, inserted and / or added; (CD3) A polypeptide that expresses cytoplasmic male sterility, consisting of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 8; (CE) Any of the following polypeptides (CE1) to (CE3): (CE1) a polypeptide consisting of the amino acid sequence of SEQ ID NO: 10; (CE2) A polypeptide that expresses cytoplasmic male sterility, consisting of the amino acid sequence of SEQ ID NO: 10 in which one or several amino acids have been deleted, substituted, inserted and / or added; (CE3) A polypeptide that expresses cytoplasmic male sterility, consisting of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 10; (CF) Any of the following polypeptides (CF1) to (CF3): (CF1) a polypeptide consisting of the amino acid sequence of SEQ ID NO: 12; (CF2) A polypeptide that expresses cytoplasmic male sterility, consisting of the amino acid sequence of SEQ ID NO: 12 in which one or several amino acids have been deleted, substituted, inserted and / or added; (CF3) A polypeptide consisting of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 12, which expresses cytoplasmic male sterility.

[0157] The (CA1) is a protein predicted to cause cytoplasmic male sterility, and is a polypeptide encoded by the nucleotide sequence of SEQ ID NO: 1. The amino acid sequence of SEQ ID NO: 2 can be obtained, for example, from seeds of the Rudbeckia plant deposited under Accession No. FERM BP-22428.

[0158] In the (CA2) amino acid sequence, the "one or several" may be within a range in which the (CA2) polypeptide expresses the cytoplasmic male sterility. The "one or several" in the (CA2) amino acid sequence may be, for example, 1 to 44, 1 to 33, 1 to 22, 1 to 11, 1 to 8, 1 to 6, 1 to 4, 1 to 3, 1 or 2, or 1 in the amino acid sequence of SEQ ID NO: 2.

[0159] The "identity" of the amino acid sequence of (CA3) may be within a range in which the polypeptide of (CA3) expresses the cytoplasmic male sterility, for example, 80% or more, 85% or more, 90% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more with respect to the amino acid sequence of SEQ ID NO: 2.

[0160] The (CB1) is a protein predicted to cause cytoplasmic male sterility, and is a polypeptide encoded by the nucleotide sequence of SEQ ID NO: 3. The amino acid sequence of SEQ ID NO: 4 can be obtained, for example, from seeds of the Rudbeckia plant deposited under Accession No. FERM BP-22428.

[0161] In the (CB2) amino acid sequence, the "one or several" may be within a range in which the (CB2) polypeptide expresses the cytoplasmic male sterility. The "one or several" in the (CB2) amino acid sequence may be, for example, 1 to 187, 1 to 140, 1 to 93, 1 to 46, 1 to 35, 1 to 28, 1 to 18, 1 to 9, 1 to 8, 1 to 6, 1 to 4, 1 to 3, 1 or 2, or 1 in the amino acid sequence of SEQ ID NO: 4.

[0162] The "identity" of the amino acid sequence of (CB3) may be within a range in which the polypeptide of (CB3) expresses the cytoplasmic male sterility, for example, and the identity of (CB3) to the amino acid sequence of SEQ ID NO: 4 is, for example, 80% or more, 85% or more, 90% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more.

[0163] The (CC1) is a protein predicted to cause cytoplasmic male sterility, and is a polypeptide encoded by the nucleotide sequence of SEQ ID NO: 5. The amino acid sequence of SEQ ID NO: 6 can be obtained, for example, from seeds of the Rudbeckia plant deposited under Accession No. FERM BP-22428.

[0164] In the amino acid sequence of (CC2), the "one or several" may be within a range in which the polypeptide of (CC2) expresses the cytoplasmic male sterility. For example, the "one or several" in (CC2) may be, for example, 1 to 86, 1 to 63, 1 to 42, 1 to 21, 1 to 16, 1 to 12, 1 to 8, 1 to 6, 1 to 4, 1 to 3, 1 or 2, or 1 in the amino acid sequence of SEQ ID NO: 6.

[0165] The "identity" of the amino acid sequence of (CC3) may be within a range that allows the polypeptide of (CC3) to express the cytoplasmic male sterility, for example, the identity of (CC3) to the amino acid sequence of SEQ ID NO: 6 is, for example, 80% or more, 85% or more, 90% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more.

[0166] The (CD1) is a protein predicted to cause cytoplasmic male sterility, and is a polypeptide encoded by the nucleotide sequence of SEQ ID NO: 7. The amino acid sequence of SEQ ID NO: 8 can be obtained, for example, from seeds of the Rudbeckia plant deposited under Accession No. FERM BP-22428.

[0167] In (CD2), the "one or several" in the amino acid sequence may be within a range in which the polypeptide of (CD2) expresses the cytoplasmic male sterility. For example, the "one or several" in (CD2) may be, for example, 1 to 194, 1 to 145, 1 to 97, 1 to 48, 1 to 38, 1 to 29, 1 to 18, 1 to 16, 1 to 12, 1 to 9, 1 to 8, 1 to 6, 1 to 4, 1 to 3, 1 or 2, or 1 in the amino acid sequence of SEQ ID NO: 8.

[0168] The "identity" of the amino acid sequence of (CD3) may be within a range in which the polypeptide of (CD3) expresses the cytoplasmic male sterility, for example, and the identity of (CD3) to the amino acid sequence of SEQ ID NO: 8 is, for example, 80% or more, 85% or more, 90% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more.

[0169] The (CE1) is a protein predicted to cause cytoplasmic male sterility, and is a polypeptide encoded by the nucleotide sequence of SEQ ID NO: 9. The amino acid sequence of SEQ ID NO: 10 can be obtained, for example, from seeds of the Rudbeckia plant deposited under Accession No. FERM BP-22428.

[0170] In the amino acid sequence of (CE2), the "one or several" may be within a range in which the polypeptide of (CE2) expresses the cytoplasmic male sterility. For example, the "one or several" in (CE2) may be, for example, 1 to 76, 1 to 57, 1 to 38, 1 to 19, 1 to 15, 1 to 12, 1 to 9, 1 to 8, 1 to 6, 1 to 4, 1 to 3, 1 or 2, or 1 in the amino acid sequence of SEQ ID NO: 10.

[0171] The "identity" of the amino acid sequence of (CE3) may be within a range in which the polypeptide of (CE3) expresses the cytoplasmic male sterility, for example, 80% or more, 85% or more, 90% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more with respect to the amino acid sequence of SEQ ID NO: 10.

[0172] The (CF1) is a protein predicted to cause cytoplasmic male sterility, and is a polypeptide encoded by the nucleotide sequence of SEQ ID NO: 11. The amino acid sequence of SEQ ID NO: 12 can be obtained, for example, from seeds of the Rudbeckia plant deposited under Accession No. FERM BP-22428.

[0173] In the amino acid sequence of (CF2), the "one or several" may be within a range in which the polypeptide of (CF2) expresses the cytoplasmic male sterility. For example, the "one or several" in (CF2) may be, for example, 1 to 31, 1 to 23, 1 to 15, 1 to 7, 1 to 6, 1 to 4, 1 to 3, 1 or 2, or 1 in the amino acid sequence of SEQ ID NO: 12.

[0174] The "identity" of the amino acid sequence of (CF3) may be within a range in which the polypeptide of (CF3) expresses the cytoplasmic male sterility, for example, 80% or more, 85% or more, 90% or more, 95% or more, 96% or more, 97% or more, 98% or more, or 99% or more with respect to the amino acid sequence of SEQ ID NO: 12.

[0175] The cytoplasmic male sterility protein of the present disclosure may comprise one or more, a plurality, or all of the polypeptides (CA) to (CF). When the cytoplasmic male sterility protein of the present disclosure comprises a plurality of polypeptides, the combination of the polypeptides can be described by replacing (ca), (cb), (cc), (cd), (ce), (cf), and polynucleotides with (CA), (CB), (CC), (CD), (CE), (CF), and polypeptides, respectively, in the description of the combination of a plurality of cytoplasmic male sterility genes in a Rudbeckia plant of the present disclosure.

[0176] The male sterility protein of the present disclosure can induce cytoplasmic male sterility, for example, by introducing it into a Rudbeckia plant. The cytoplasmic male sterility protein of the present disclosure can also be referred to as, for example, a recombinant protein.

[0177] The cytoplasmic male sterility protein can be synthesized (produced), for example, using the transformant described below.

[0178] <Expression vector> In another aspect, the present disclosure provides an expression vector capable of expressing a cytoplasmic male sterility protein. The expression vector (vector) of the present disclosure comprises a cytoplasmic male sterility gene of the present disclosure. The expression vector of the present disclosure can be suitably used to synthesize (produce) a Rudbeckia plant exhibiting cytoplasmic male sterility of the present disclosure by genetic engineering techniques.

[0179] The expression vector may contain the cytoplasmic male sterility gene so as to be capable of expressing the cytoplasmic male sterility protein encoded by the cytoplasmic male sterility gene, and other configurations and conditions are not particularly limited. In the expression vector of the present disclosure, the cytoplasmic male sterility gene of the present disclosure may be said to be functionally linked.

[0180] The expression vector may contain one or more, a plurality of, or all of the polynucleotides (ca) to (cf). When the expression vector contains a plurality of polynucleotides, the combination of the polynucleotides can be determined by the same explanation as for the combination of a plurality of cytoplasmic male sterility genes in Rudbeckia plants of the present disclosure.

[0181] The expression vector can be prepared, for example, by inserting a polynucleotide encoding a cytoplasmic male sterility protein, i.e., the cytoplasmic male sterility gene, into a backbone vector (hereinafter also referred to as a "basic vector"). The type of vector is not particularly limited and can be appropriately determined depending on, for example, the type of host or cell.

[0182] Examples of the host include non-human hosts such as microorganisms, animal cells, plant cells, insect cells, or cultured cells thereof, isolated human cells or cultured cells thereof, and mammalian cells.

[0183] Examples of the vector (basic vector) include viral vectors and non-viral vectors. When the heat shock method is used to transform a host, the vector may be, for example, a binary vector. Examples of the vector include pETDuet-1, pQE-80L, and pUCP26Km. When bacteria such as Escherichia coli are transformed, examples of the vector include pETDuet-1 vector (Novagen), pQE-80L (QIAGEN), pBR322, pB325, pAT153, and pUC8. When yeast is transformed, examples of the vector include pYepSec1, pMFa, and pYES2. When insect cells are transformed, examples of the vector include pAc and pVL. When mammalian cells are transformed, examples of the vector include pCDM8 and pMT2PC. When transforming the plant cells, examples of the vector include pBI121 and pBI101.

[0184] The expression vector preferably contains a regulatory sequence that regulates the expression of the polynucleotide encoding the cytoplasmic male sterility protein (cytoplasmic male sterility gene) and the expression of the cytoplasmic male sterility protein encoded by the cytoplasmic male sterility gene. Examples of the regulatory sequence include a promoter, a terminator, an enhancer, a polyadenylation signal sequence, and an origin of replication (ori). The location of the regulatory sequence in the expression vector of the present disclosure is not particularly limited. The regulatory sequence in the expression vector of the present disclosure may be located, for example, so long as it is capable of functionally regulating the expression of the polynucleotide encoding the cytoplasmic male sterility protein and the expression of the male sterility protein encoded thereby, and can be located according to known methods. The regulatory sequence may be, for example, a sequence already present in the basic vector, or the regulatory sequence may be inserted into the basic vector, or the regulatory sequence present in the basic vector may be replaced with another regulatory sequence.

[0185] The expression vector may further comprise, for example, a coding sequence for a selection marker, such as a drug resistance marker, a fluorescent protein marker, an enzyme marker, or a cell surface receptor marker.

[0186] Insertion of DNA, insertion of the regulatory sequence, and / or insertion of the coding sequence of the selection marker into the expression vector may be carried out, for example, by a method using restriction enzymes and ligase, or by using a commercially available kit, etc.

[0187] <Transformants> In another aspect, the present disclosure provides a transformant capable of expressing a cytoplasmic male sterility protein. The transformant of the present disclosure comprises a nucleic acid or an expression vector of the present disclosure. The transformant of the present disclosure can also be said to comprise an exogenous cytoplasmic male sterility gene. Furthermore, the transformant of the present disclosure can be suitably used for producing a Rudbeckia plant exhibiting cytoplasmic male sterility or for synthesizing (producing) a cytoplasmic male sterility protein.

[0188] In the transformant of the present disclosure, the cytoplasmic male sterility gene of the present disclosure is present as an exogenous molecule, and therefore, the transformant of the present disclosure can be produced, for example, by introducing the cytoplasmic male sterility gene into the host.

[0189] The method for introducing the cytoplasmic male sterility gene is not particularly limited and can be performed by a known method. The cytoplasmic male sterility gene may be introduced, for example, using the expression vector of the present disclosure. The introduction method can be appropriately selected depending on, for example, the type of host. Examples of the introduction method include introduction using a gene gun such as a particle gun, the calcium phosphate method, the polyethylene glycol method, lipofection using liposomes, electroporation, ultrasonic nucleic acid transfer, DEAE-dextran, direct injection using glass microtubes, the hydrodynamic method, the cationic liposome method, a method using an introduction adjuvant, and an Agrobacterium-mediated method. Examples of the liposome include lipofectamine and cationic liposomes, and examples of the introduction adjuvant include atelocollagen, nanoparticles, and polymers. When the host is a plant cell, the Agrobacterium-mediated introduction method is preferred. When the host is a plant cell, the target into which the cytoplasmic male sterility gene is introduced may be, for example, a plant cell, a callus, a plant tissue, or an individual plant.

[0190] The method for introducing the cytoplasmic male sterility gene can be carried out, for example, by combining a genome editing technique using homologous recombination; ZFN, TALEN, CRISPR-CAS9, CRISPR-CPF1, or the like, with a donor gene (cytoplasmic male sterility gene). Introduction of the cytoplasmic male sterility gene using the genome editing technique can be carried out, for example, by introducing proteins and nucleic acids constituting the genome editing technique, or vectors encoding these, and a polynucleotide encoding the donor gene. The protein can be, for example, CRISPR (Clustered Regularly Interspaced Short Palindromic Recombination). Repeats) enzymes, specific examples of which include Cas1, Cas1B, Cas2, Cas3, Cas4, Cas5, Cas6, Cas7, Cas8, Cas9, Cas10, Csy1, Csy2, Csy3, Cse1, Cse2, Csc1, Csc2, Csa5, Csn2, Csm2, Csm3, Csm4, Csm5, Csm6, Cmr1, Cmr3, Cmr4, Cmr5, Cmr6, Csb1, Csb2, Csb3, Csx17, Csx14, Csx10, Csx16, CsaX, Csx3, Csx1, Csx15, Csf1, Csf2, Csf3, and Csf4. The nucleic acid may be, for example, a crRNA or a tracrRNA, or a single-stranded nucleic acid formed by linking these together via a linker. In this case, the nucleic acid is designed such that the base sequence that anneals to the target sequence in the crRNA is complementary to the base sequence encoding each gene. The nucleic acid may be used alone or in combination of two or more types.

[0191] The method for introducing the polynucleotide is not particularly limited and can be, for example, a method used in RNA interference, antisense RNA, genome editing technology, etc. An expression cassette such as an expression vector containing the polynucleotide can be introduced into a target Rudbeckia plant by, for example, the polyethylene glycol method, electroporation, Agrobacterium-mediated method, particle gun method, etc. The target Rudbeckia plant may be, for example, a plant cell, callus, plant tissue, or an individual plant.

[0192] In the introduction step, the cytoplasmic male sterility gene may be introduced into, for example, the cytoplasm, mitochondria, or mitochondrial genome.

[0193] <How to grant> In another aspect, the present disclosure provides a method for imparting cytoplasmic male sterility to a Rudbeckia plant. As described above, the method for imparting cytoplasmic male sterility to a Rudbeckia plant of the present disclosure includes the step of introducing, into a target Rudbeckia plant, at least one polynucleotide selected from the group consisting of the following (ca) to (ce) and (cf) as a cytoplasmic male sterility gene (introduction step):

[0194] (ca) any one of the following polynucleotides (ca1) to (ca7): (ca1) a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1; (ca2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (ca1); (ca3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (ca1), and that causes cytoplasmic male sterility; (ca4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (ca1); (ca5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 2; (ca6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 2 in which one or several amino acids have been deleted, substituted, inserted and / or added; (ca7) a polynucleotide that expresses cytoplasmic male sterility and that consists of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 2; (cb) any one of the following polynucleotides (cb1) to (cb7): (cb1) a polynucleotide consisting of the base sequence of SEQ ID NO: 3; (cb2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cb1); (cb3) a polynucleotide that expresses cytoplasmic male sterility and that consists of a base sequence having 80% or more identity to the base sequence of (cb1); (cb4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cb1); (cb5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 4; (cb6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 4 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cb7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 4; (cc) any one of the following polynucleotides (cc1) to (cc7): (cc1) a polynucleotide consisting of the base sequence of SEQ ID NO: 5; (cc2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cc1); (cc3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (cc1), and that expresses cytoplasmic male sterility; (cc4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to the polynucleotide having the nucleotide sequence of (cc1); (cc5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 6; (cc6) A polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 6 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cc7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 6; (cd) any one of the following polynucleotides (cd1) to (cd7): (cd1) a polynucleotide consisting of the base sequence of SEQ ID NO: 7; (cd2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cd1); (cd3) a polynucleotide consisting of a base sequence having 80% or more identity to the base sequence of (cd1), which expresses cytoplasmic male sterility; (cd4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cd1); (cd5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 8; (cd6) A polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 8 in which one or several amino acids have been deleted, substituted, inserted and / or added; (cd7) a polynucleotide that expresses cytoplasmic male sterility and consists of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 8; (ce) any one of the following polynucleotides (ce1) to (ce7): (ce1) a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 9; (ce2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (ce1); (ce3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (ce1), and that causes cytoplasmic male sterility; (ce4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the nucleotide sequence of (ce1); (ce5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 10; (ce6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 10 in which one or several amino acids have been deleted, substituted, inserted and / or added; (ce7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 10; (cf) any one of the following polynucleotides (cf1) to (cf7): (cf1) a polynucleotide consisting of the base sequence of SEQ ID NO: 11; (cf2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cf1); (cf3) a polynucleotide that expresses cytoplasmic male sterility and that consists of a base sequence having 80% or more identity to the base sequence of (cf1); (cf4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to the polynucleotide having the nucleotide sequence of (cf1); (cf5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 12; (cf6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 12 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cf7) A polynucleotide consisting of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 12, which polynucleotide causes expression of cytoplasmic male sterility.

[0195] The method of the present disclosure is characterized by introducing at least one polynucleotide selected from the group consisting of (ca) to (ce) and (cf) as a cytoplasmic male sterility gene into a target Rudbeckia plant, and other steps and conditions are not particularly limited. According to the method of the present disclosure, cytoplasmic male sterility can be conferred to a Rudbeckia plant.

[0196] In the introduction step, the method for introducing the cytoplasmic male sterility gene into the target Rudbeckia plant can be carried out in the same manner as the method for introducing the cytoplasmic male sterility gene in the production of the transformant, for example.

[0197] In the introduction step, the polynucleotide to be introduced may include one or more, a plurality of, or all of the polynucleotides (ca) to (cf). When the polynucleotide to be introduced includes a plurality of polynucleotides, the combination of the polynucleotides can be determined by referring to the description of the combination of a plurality of cytoplasmic male sterility genes in Rudbeckia plants of the present disclosure.

[0198] <Second production method> In another aspect, the present disclosure provides a method for producing a Rudbeckia plant exhibiting cytoplasmic male sterility. As described above, the method for producing a cytoplasmic male sterile Rudbeckia plant of the present disclosure includes a step of conferring cytoplasmic male sterility to a target Rudbeckia plant, and the conferring step is carried out by the method for conferring cytoplasmic male sterility in a Rudbeckia plant of the present disclosure. A second production method of the present disclosure is characterized in that the conferring step is carried out by the method for conferring cytoplasmic male sterility in a Rudbeckia plant of the present disclosure, and other steps and conditions are not particularly limited. According to the second production method of the present disclosure, a cytoplasmic male sterile Rudbeckia plant can be produced.

[0199] <Screening method> In another aspect, the present disclosure provides a method for screening for Rudbeckia plants exhibiting cytoplasmic male sterility. The method for screening for cytoplasmic male sterile Rudbeckia plants of the present disclosure includes the step of selecting, from test Rudbeckia plants, test Rudbeckia plants that contain, as a cytoplasmic male sterility gene, at least one polynucleotide selected from the group consisting of (ca) to (ce) and (cf) below, as cytoplasmic male sterility genes.

[0200] (ca) any one of the following polynucleotides (ca1) to (ca7): (ca1) a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1; (ca2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (ca1); (ca3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (ca1), and that causes cytoplasmic male sterility; (ca4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (ca1); (ca5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 2; (ca6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 2 in which one or several amino acids have been deleted, substituted, inserted and / or added; (ca7) a polynucleotide that expresses cytoplasmic male sterility and that consists of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 2; (cb) any one of the following polynucleotides (cb1) to (cb7): (cb1) a polynucleotide consisting of the base sequence of SEQ ID NO: 3; (cb2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cb1); (cb3) a polynucleotide that expresses cytoplasmic male sterility and that consists of a base sequence having 80% or more identity to the base sequence of (cb1); (cb4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cb1); (cb5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 4; (cb6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 4 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cb7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 4; (cc) any one of the following polynucleotides (cc1) to (cc7): (cc1) a polynucleotide consisting of the base sequence of SEQ ID NO: 5; (cc2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cc1); (cc3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (cc1), and that expresses cytoplasmic male sterility; (cc4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to the polynucleotide having the nucleotide sequence of (cc1); (cc5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 6; (cc6) A polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 6 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cc7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 6; (cd) any one of the following polynucleotides (cd1) to (cd7): (cd1) a polynucleotide consisting of the base sequence of SEQ ID NO: 7; (cd2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cd1); (cd3) a polynucleotide consisting of a base sequence having 80% or more identity to the base sequence of (cd1), which expresses cytoplasmic male sterility; (cd4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cd1); (cd5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 8; (cd6) A polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 8 in which one or several amino acids have been deleted, substituted, inserted and / or added; (cd7) a polynucleotide that expresses cytoplasmic male sterility and consists of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 8; (ce) any one of the following polynucleotides (ce1) to (ce7): (ce1) a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 9; (ce2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (ce1); (ce3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (ce1), and that causes cytoplasmic male sterility; (ce4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the nucleotide sequence of (ce1); (ce5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 10; (ce6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 10 in which one or several amino acids have been deleted, substituted, inserted and / or added; (ce7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 10; (cf) any one of the following polynucleotides (cf1) to (cf7): (cf1) a polynucleotide consisting of the base sequence of SEQ ID NO: 11; (cf2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cf1); (cf3) a polynucleotide that expresses cytoplasmic male sterility and that consists of a base sequence having 80% or more identity to the base sequence of (cf1); (cf4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to the polynucleotide having the nucleotide sequence of (cf1); (cf5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 12; (cf6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 12 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cf7) A polynucleotide consisting of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 12, which polynucleotide causes expression of cytoplasmic male sterility.

[0201] The screening method of the present disclosure is characterized in that the cytoplasmic male gene is used as an indicator in the selection step, and other steps and conditions are not particularly limited. According to the screening method of the present disclosure, a cytoplasmic male sterile Rudbeckia plant can be screened.

[0202] In the screening method of the present disclosure, the selection step can be performed using the explanation for the indirect selection (selection using indirect evaluation) in the step (x) or the step (x1).

[0203] <Third production method> In another aspect, the present disclosure provides a method for producing a Rudbeckia plant exhibiting cytoplasmic male sterility. As described above, the method for producing a cytoplasmic male sterile Rudbeckia plant of the present disclosure includes a screening step of screening test Rudbeckia plants for test Rudbeckia plants containing a cytoplasmic male sterility gene, and the screening step is carried out by the screening method of the present disclosure. A third production method of the present disclosure is characterized in that the screening step is carried out by the screening method of the present disclosure, and other steps and conditions are not particularly limited.

[0204] <The second Rudbeckia plant> In another aspect, the present disclosure provides a Rudbeckia plant that exhibits cytoplasmic male sterility. The cytoplasmic male sterile Rudbeckia plant of the present disclosure (hereinafter also referred to as the "second Rudbeckia plant") is obtained by the first production method, the second production method, or the third production method of the present disclosure. The second Rudbeckia plant of the present disclosure is characterized in that it is obtained by the first production method, the second production method, or the third production method of the present disclosure, and other configurations and conditions are not particularly limited.

[0205] <Detection method> In another aspect, the present disclosure provides a method for detecting cytoplasmic male sterility in a Rudbeckia plant. As described above, the method for detecting cytoplasmic male sterility in a Rudbeckia plant of the present disclosure includes a detection step of detecting, in a test Rudbeckia plant, at least one polynucleotide selected from the group consisting of the following (ca) to (ce) and (cf) as a cytoplasmic male sterility gene:

[0206] (ca) any one of the following polynucleotides (ca1) to (ca7): (ca1) a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1; (ca2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (ca1); (ca3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (ca1), and that causes cytoplasmic male sterility; (ca4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (ca1); (ca5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 2; (ca6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 2 in which one or several amino acids have been deleted, substituted, inserted and / or added; (ca7) a polynucleotide that expresses cytoplasmic male sterility and that consists of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 2; (cb) any one of the following polynucleotides (cb1) to (cb7): (cb1) a polynucleotide consisting of the base sequence of SEQ ID NO: 3; (cb2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cb1); (cb3) a polynucleotide that expresses cytoplasmic male sterility and that consists of a base sequence having 80% or more identity to the base sequence of (cb1); (cb4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cb1); (cb5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 4; (cb6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 4 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cb7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 4; (cc) any one of the following polynucleotides (cc1) to (cc7): (cc1) a polynucleotide consisting of the base sequence of SEQ ID NO: 5; (cc2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cc1); (cc3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (cc1), and that expresses cytoplasmic male sterility; (cc4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to the polynucleotide having the nucleotide sequence of (cc1); (cc5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 6; (cc6) A polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 6 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cc7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 6; (cd) any one of the following polynucleotides (cd1) to (cd7): (cd1) a polynucleotide consisting of the base sequence of SEQ ID NO: 7; (cd2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cd1); (cd3) a polynucleotide consisting of a base sequence having 80% or more identity to the base sequence of (cd1), which expresses cytoplasmic male sterility; (cd4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cd1); (cd5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 8; (cd6) A polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 8 in which one or several amino acids have been deleted, substituted, inserted and / or added; (cd7) a polynucleotide that expresses cytoplasmic male sterility and consists of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 8; (ce) any one of the following polynucleotides (ce1) to (ce7): (ce1) a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 9; (ce2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (ce1); (ce3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (ce1), and that causes cytoplasmic male sterility; (ce4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the nucleotide sequence of (ce1); (ce5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 10; (ce6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 10 in which one or several amino acids have been deleted, substituted, inserted and / or added; (ce7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 10; (cf) any one of the following polynucleotides (cf1) to (cf7): (cf1) a polynucleotide consisting of the base sequence of SEQ ID NO: 11; (cf2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cf1); (cf3) a polynucleotide that expresses cytoplasmic male sterility and that consists of a base sequence having 80% or more identity to the base sequence of (cf1); (cf4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to the polynucleotide having the nucleotide sequence of (cf1); (cf5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 12; (cf6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 12 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cf7) A polynucleotide consisting of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 12, which polynucleotide causes expression of cytoplasmic male sterility.

[0207] The detection method of the present disclosure is characterized in that the cytoplasmic male sterility gene is detected in the detection step, and other steps and conditions are not particularly limited. According to the detection method of the present disclosure, it is possible to detect whether a test Rudbeckia plant has cytoplasmic male sterility.

[0208] The detection method of the present disclosure includes, for example, a detection step of detecting a cytoplasmic male sterility gene in a test Rudbeckia plant. The detection step can be performed using the same explanation as for step (x) in the selection step of the first production method of the present disclosure or the indirect selection (selection using indirect evaluation) in step (x1).

[0209] In the selection step, the location of the cytoplasmic male sterility gene may be detected. Examples of the location include the nucleus, the nuclear genome, the cytoplasm, the mitochondria, and the mitochondrial genome.

[0210] In the selection step, the sample for detecting the cytoplasmic male sterility gene may be, for example, a sample containing the nucleus, nuclear genome, cytoplasm, mitochondria, and / or mitochondrial genome. Each sample can be prepared from the test Rudbeckia plant or a part thereof by a conventional method in the technical field of the present application. The sample is preferably a sample containing the cytoplasm, mitochondria, and / or mitochondrial genome.

[0211] <Deposited strain> An example of a Rudbeckia plant of the present disclosure is a Rudbeckia plant identified (deposited) under accession number FERM BP-22428 or its progeny. Hereinafter, accession number FERM BP-22428 will also be referred to as Rudbeckia cultivar Takii 22. Information regarding the deposit of each cultivar is provided below.

[0212] Type of deposit: International deposit Name of depository institution: National Institute of Technology and Evaluation, Patent Organism Depositary Center Address: Room 120, 2-5-8 Kazusa Kamatari, Kisarazu City, Chiba Prefecture, 292-0818, Japan Accession number: Accession number FERM BP-22428 Identification mark: Takii 22 Received: September 14, 2021

[0213] In this disclosure, a plant having "essentially all physiological and morphological characteristics of the deposited line" means a plant that has the major traits of the deposited line when grown in the same environment.

[0214] <Progeny lineage> The Rudbeckia plant of the present disclosure may be a progeny line of the deposited line. The progeny line may be an individual plant of the progeny line, a part of an individual plant of the progeny line, or a seed of the progeny line.

[0215] In the present disclosure, a "progeny line" or "progeny Rudbeckia plant" (hereinafter collectively referred to as "progeny line") refers to a plant obtained from a Rudbeckia plant of a deposited line or its progeny line. In the present disclosure, the progeny line may be a plant obtained by crossing the deposited line with another deposited line or another Rudbeckia plant, or by crossing the deposited line with a wild Rudbeckia plant. The progeny line may be obtained, obtainable, or derived directly or indirectly by cross-pollinating the deposited line or its progeny line, or may be derived from a parent line obtained from the deposited line using traditional breeding methods such as cross-pollination. Examples of the progeny line include a first-generation hybrid F1 (hybrid first generation line, F1 hybrid), a backcross progeny line, and the like. In obtaining the progeny line, the deposited line exhibits cytoplasmic male sterility, as described above. Therefore, the deposited line is used as a female parent.

[0216] In this disclosure, "cross" refers to the crossing of two parent lines. The cross may be "cross-pollination" or "self-pollination." In this disclosure, when the cross is used in conjunction with a plant that exhibits cytoplasmic male sterility, the cross refers to cross-pollination. The cross-pollination refers to fertilization by the union of two gametes from different plants. The self-pollination refers to the transfer of pollen from an anther to the stigma of the same plant. The self-pollination can also be referred to as, for example, selfing. The cross may include backcrossing, which is a traditional breeding method.

[0217] The "backcrossing" technique is a traditional breeding technique in which breeders repeatedly backcross a progeny line of a hybrid with one of the parent lines to introduce a trait into a plant or variety. A plant containing the trait to be introduced can be referred to as, for example, a donor plant (donor parent). The plant into which the trait is introduced can be referred to as, for example, a recurrent parent. Backcrossing can be performed by crossing a donor plant with the recurrent parent, thereby obtaining a first-generation hybrid F1 (first-generation hybrid line, F1 hybrid). Next, the progeny line containing the trait is crossed with the recurrent parent. Then, by backcrossing for several generations, the trait of the donor plant can be introduced into the recurrent parent. The Rudbeckia plant of the present disclosure may be used as the donor plant.

[0218] In the present disclosure, the progeny line may be regenerated from a cell culture or tissue culture, protoplast, or part of a plant derived from the deposited line, or may be obtained by self-pollination of the deposited line, or may be obtained by producing seeds from a plant of the deposited line.

[0219] In the present disclosure, the term "regeneration" refers to the development or vegetative propagation of plants from cell culture, tissue culture, or protoplasts.

[0220] The "tissue culture" or "cell culture" may be a composition containing isolated cells of the same or different types, or may be a collection of cells organized into plant parts. Methods for tissue culture of various tissues of Rudbeckia plants and for regenerating plants from said tissue cultures are well known; see, for example, Reference 1 below. Reference 1: P. Szarvas et.al., “Biotechnology of annual plants flower: Micropropagation of Rudbeckia sp”, Acta Horticulturae, 2006, vol. 725, pages 527-548

[0221] The cytoplasmic male sterility in the deposited line is a dominant hereditary trait, and by crossing with a male parent, the progeny line inherits the cytoplasm of the cytoplasmic male sterile female parent, and therefore the progeny line has cytoplasmic male sterility.

[0222] The progeny line may have desired traits, for example, "essentially all of the physiological and morphological characteristics of the deposited line" when grown under the same cultivation conditions.

[0223] The progeny line may include cells containing at least one set of chromosomes derived from the corresponding deposited line. The progeny line may, for example, have at least 6.25%, 12.5%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% of its alleles derived from the corresponding deposited line. That is, the progeny line may have at least about 6.25%, 12.5%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% genetic complement with the corresponding deposited line.

[0224] An "allele" is any one or more genes, all of which are associated with a characteristic or trait in a Rudbeckia plant. In a diploid cell or organism, the pair of alleles of a given gene occupy corresponding loci on a pair of homologous chromosomes.

[0225] The genetic complementarity can be calculated, for example, by decoding molecular markers or nucleotide sequences, comparing them with the molecular markers or nucleotide sequences of the deposited line, and calculating the percentage of identity. Examples of molecular markers include single nucleotide polymorphism (SNP) markers, amplified fragment length polymorphism (AFLP) markers, restriction fragment length polymorphism (RFLP) markers, microsatellite markers, sequence-characterized amplified region (SCAR) markers, and cleaved amplified polymorphic sequence (CAPS) markers. Genome analysis methods using molecular markers are well known and widely published (e.g., References 2 and 3 below). The nucleotide sequence can be determined, for example, by extracting chromosomes from the progeny line and sequencing the chromosomes. The proportion of alleles derived from the deposited line and the proportion of genetic complementarity can be estimated, for example, by the number of crosses. In this case, the proportion can be estimated from the number of crosses from the deposited line. As a specific example, when the number of crosses from the deposited strain is n, the ratio is, for example, (1 / 2) n It can be estimated as ×100%. Reference 2: Sinchan Adhikari et.al, “Application of molecular markers in plant genome analysis: a review”, The Nucleus, 2017, Volume 60, Issue 3, pp. 283-297 Reference 3: Elcio P. Guimaraes et.al., “MARKER-ASSISTED SELECTION Current status and future perspectives in crops, livestock, forestry and fish”, 2007, Springer, 29-49

[0226] The proportion of alleles and genetic complementarity derived from the deposited line is preferably, for example, the average of the proportions of multiple progeny lines, where the number is, for example, the number of individuals that can be statistically analyzed, specifically, 200 or more individuals, preferably 200 to 1000 individuals.

[0227] The progeny line may have SNPs derived from the deposited line. For example, the progeny line may have at least 6.25%, 12.5%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% of its SNPs derived from the deposited line. That is, the progeny line may be at least about 6.25%, 12.5%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% SNP-identical to the deposited line. In the present disclosure, if, for example, 50%, 55%, 60%, 65%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more of the SNPs of a target Rudbeckia plant match those of the deposited line, the target Rudbeckia plant can be determined (identified, estimated, identified, or differentiated) to be a descendant line of the deposited line.

[0228] The progeny line may have, for example, a mutation or an introduced gene. In this case, the progeny line has, for example, one or more modified traits. The progeny line can be produced, for example, by introducing a mutation or an introduced gene into the deposited line or its progeny line. The mutation may be artificially introduced or naturally introduced. The mutation may be, for example, a chemically induced mutation or a radiation-induced mutation. The mutation may also be introduced by, for example, a molecular biological technique or a genome editing technique (see, for example, Reference 4 below). The introduced gene can be introduced, for example, by a method using Agrobacterium tumefaciens (see, for example, Reference 4 below). Reference 4: Yanfei Mao et.al., “Gene editing in plants: progress and challenges”, National Science Review, 2019, vol. 6, pp. 421-437

[0229] The term "transgene" refers to a desired gene introduced into the genome of a plant, for example, by genetic engineering techniques or traditional breeding methods. The transgene may be derived from the same species or a different species. The transgene may be a gene containing the same nucleotide sequence as that of the species from which it originates, or a gene containing a different nucleotide sequence. In the latter case, the different nucleotide sequence can be prepared, for example, by codon optimization, addition of a transcriptional regulator such as a promoter, or the like, to the same nucleotide sequence. The transgene may include a translated region and an untranslated region.

[0230] <Haploid and doubled haploid plants> The Rudbeckia plants of the present disclosure may be haploid and / or doubled haploid plants obtained, obtainable, or derived from the deposited line. The haploid and / or doubled haploid plants of the deposited line may be used in methods for producing parent lines of the deposited line. In one embodiment, the present disclosure may provide haploid and / or doubled haploid plants, plant parts of haploid and / or doubled haploid plants, or seeds of haploid and / or doubled haploid plants.

[0231] The doubled haploid plant can be produced by doubling the chromosomes in a haploid plant or cell. Specifically, haploid cytoplasm is cultured under specific conditions to form plantlets with 1n chromosomes. The plantlets are then treated with a chemical substance such as colchicine to double the chromosomes. As a result, the cells of the plantlets have 2n chromosomes (doubled haploid). The doubled haploid plant and its progeny can then be obtained by growing the plantlets after the treatment.

[0232] <Method of producing Rudbeckia plants> As described above, the method for producing a Rudbeckia plant of the present disclosure includes a hybridization step of hybridizing a first Rudbeckia plant with a second Rudbeckia plant, the first Rudbeckia plant being the Rudbeckia plant of the present disclosure. The method for producing a Rudbeckia plant of the present disclosure is characterized in that the Rudbeckia plant of the present disclosure is used as at least one of the parents in the hybridization step, and other steps and conditions are not particularly limited.

[0233] Furthermore, the method for producing a Rudbeckia plant of the present disclosure includes a hybridization step of cross-pollinating (cross-pollinating) the Rudbeckia plants of the present disclosure. The production method of the present disclosure is characterized by cross-pollinating the Rudbeckia plants of the present disclosure, and other steps and conditions are not particularly limited.

[0234] According to the production method of the present disclosure, progeny lines of the deposited line can be produced. The production method of the present disclosure can be applied to the description of Rudbeckia plants of the present disclosure.

[0235] In the present disclosure, the cross between the first Rudbeckia plant (first parent line) and the second Rudbeckia plant (second parent line) is a cross (cross-pollination) between different individuals.

[0236] In the present disclosure, the first parent line is the Rudbeckia plant of the present disclosure, for example, the Rudbeckia plant deposited under the aforementioned accession number FERM BP-22428 or a progeny line thereof.

[0237] The second parent line is not particularly limited, and any Rudbeckia plant can be used. The second parent line may be, for example, a Rudbeckia plant of the same taxonomic species as the first parent line, or a Rudbeckia plant of a different species. The second parent line may be, for example, a male-fertile Rudbeckia plant.

[0238] The production method of the present disclosure may further include, for example, a growing step of cultivating the progeny lines obtained in the hybridization step after the hybridization step. Examples of growing conditions in the growing step include general growing conditions for Rudbeckia plants.

[0239] The Rudbeckia plant of the present disclosure can be obtained, for example, by the production method of the present disclosure.

[0240] <Method for producing Rudbeckia seeds> The present disclosure provides a method for producing Rudbeckia seeds. The method for producing Rudbeckia seeds of the present disclosure includes crossing a Rudbeckia plant of the deposited line with another Rudbeckia plant, and optionally collecting (collecting or harvesting) the resulting seeds. The seed production method of the present disclosure may also provide a plant, plant part, or seed by growing the seeds of the Rudbeckia plant.

[0241] The seed production method of the present disclosure may be a method for producing seeds derived from the deposited line. In this case, the seed production method of the present disclosure may include the step of (a) crossing a plant of the deposited line with another Rudbeckia plant to produce seeds. The seed production method of the present disclosure may further include the steps of (b) cultivating a Rudbeckia plant from the seed of step (a) to produce a Rudbeckia plant derived from the deposited line, and (c) crossing the Rudbeckia plant of step (b) with another Rudbeckia plant to produce an additional Rudbeckia plant derived from the deposited line. Furthermore, the seed production method of the present disclosure may include (d) optionally repeating steps (b) and (c) one or more times to produce an additional Rudbeckia plant derived from the deposited line. In this case, the additional Rudbeckia plant obtained in step (c) can be used as the Rudbeckia plant cultivated in step (b) from the seed of step (a). The "one or more times" means, for example, 1 to 10 times, 3 to 7 times, or 3 to 5 times. The seed production method of the present disclosure may further include a step of collecting or harvesting the seeds. The seed production method of the present disclosure may provide seeds produced by the above method, and plants or plant parts obtained by growing the seeds.

[0242] The seed production method of the present disclosure may further include (e) identifying a progeny line having cytoplasmic male sterility in the production of a Rudbeckia plant in step (b), the creation of an additional Rudbeckia plant in step (c), or the creation of a further Rudbeckia plant derived from the deposited line obtained in step (d). In this case, the seed production method of the present disclosure preferably uses the progeny line having cytoplasmic male sterility as the Rudbeckia plant to be hybridized in the subsequent steps. The "identification" can also be referred to as, for example, identification, appraisal, identification, selection, or culling. The cytoplasmic male sterility of the progeny line is preferably the same as that of the deposited line from which it was derived, and the above explanation can be applied.

[0243] <Method for Producing Hybrid Rudbeckia Plants> The present disclosure provides methods for producing hybrid (hybrid) Rudbeckia plants. The methods for producing hybrid plants of the present disclosure include crossing a Rudbeckia plant of the present disclosure with another Rudbeckia plant. The methods for producing hybrid plants of the present disclosure may also include collecting or harvesting seeds resulting from the hybridization. The methods for producing hybrid plants of the present disclosure may also provide seeds and hybrid plants or parts of hybrid plant individuals produced by the above methods.

[0244] <Method of introducing new traits> The present disclosure provides a method for introducing at least one new characteristic or trait (hereinafter collectively referred to as "trait") into the deposited line. The disclosed method for introducing a trait can also be referred to, for example, as a method for producing a Rudbeckia plant into which a new trait has been introduced. The disclosed method for introducing a trait includes, for example, (a) crossing a plant of the deposited line with a Rudbeckia plant containing at least one new trait to produce a progeny line; and (b) selecting the progeny line containing at least one new trait. The disclosed method for introducing a trait includes, for example, (c) crossing the progeny line with a Rudbeckia plant having essentially all of the physiological and morphological characteristics of the deposited line with respect to traits other than cytoplasmic male sterility, to produce backcross progeny seeds. The disclosed method for introducing a trait includes, for example, (d) selecting backcross progeny having at least one new trait and, optionally, cytoplasmic male sterility. In steps (b) and (d), the selection of progeny lines having a new trait may be carried out, for example, by detecting the trait, or by detecting a gene or molecular marker associated with the trait. The new trait may be, for example, resistance to pathogens.

[0245] In step (b), it is preferable to select a Rudbeckia plant that contains at least one new trait and, optionally, exhibits cytoplasmic male sterility. As described above, when a Rudbeckia plant having cytoplasmic male sterility is used as a female parent, the resulting progeny lines exhibit cytoplasmic male sterility. Therefore, in selecting the progeny lines, the presence or absence of cytoplasmic male sterility may or may not be considered.

[0246] In step (d), a progeny line may be selected that has at least one new trait and, optionally, cytoplasmic male sterility, and that has essentially all of the physiological and morphological characteristics of the deposited line in step (a).

[0247] The trait introduction method of the present disclosure may optionally (e) repeat steps (c) and (d) one or more times to produce a Rudbeckia plant containing at least one new trait. In this case, the progeny line in step (c) of the trait introduction method of the present disclosure may use the backcross progeny selected in step (d). The Rudbeckia plant obtained or obtainable in step (e) may exhibit cytoplasmic male sterility and may further have essentially all of the physiological and morphological characteristics of the deposited line in step (a). The "one or more times" may be, for example, 1 to 10 times, 3 to 7 times, or 3 to 5 times. The trait introduction method of the present disclosure may also include a step of collecting or harvesting seeds. The trait introduction method of the present disclosure may also provide seeds produced by the above method, and plants or plant parts obtained by growing the seeds.

[0248] <Method for introducing transgene> The present disclosure provides a method for producing a plant derived from a deposited line that includes at least one new characteristic or trait. The method for introducing a transgene disclosed herein can also be referred to as a method for producing a Rudbeckia plant into which a new trait has been introduced.

[0249] The method for introducing a transgene disclosed herein may include, for example, introducing a mutation or transgene that confers at least one new trait into a plant of a deposited line or a progeny line. The introduction of the mutation or transgene can be carried out, for example, in the same manner as the introduction of a mutation or transgene into the progeny line. The Rudbeckia plant obtained or obtainable by the introduction step may exhibit cytoplasmic male sterility and may further possess essentially all of the physiological and morphological characteristics of the deposited line. The method for introducing a transgene disclosed herein may also include a step of collecting or harvesting seeds. The method for introducing a transgene disclosed herein may also provide seeds produced by the above method, as well as plants or plant parts obtained by growing the seeds. The new trait may be, for example, resistance to pathogens.

[0250] <Regeneration and regeneration method of Rudbeckia plants> The present disclosure provides Rudbeckia plants (hereinafter "regenerants") regenerated from cell cultures, tissue cultures, or protoplasts of the deposited line. The present disclosure may also provide cell or tissue cultures of regenerable cells or protoplasts derived from the Rudbeckia plant of the deposited line. The cells, tissues, or protoplasts may be derived from tissues including leaves, embryos, cotyledons, hypocotyls, meristematic cells, roots, root tips, anthers, flowers, seeds, or stems.

[0251] The present disclosure provides a method for propagating or multiplying a Rudbeckia plant of a deposited line. Propagation of the Rudbeckia plant of the deposited line may be vegetative propagation of the Rudbeckia plant of the deposited line. In this case, the method for regenerating a Rudbeckia plant of the present disclosure may include, for example, (a) collecting tissue capable of propagation from the plant of the deposited line; (b) culturing the tissue to obtain a propagated shoot; and (c) rooting the propagated shoot to obtain a rooted plantlet. The method for regenerating a Rudbeckia plant of the present disclosure may further include, optionally, (d) growing a plant from the rooted plantlet. For details of the vegetative propagation method, see, for example, Reference 5 below. The regeneration method of the present disclosure may provide, for example, a plantlet, plant, or plant part regenerated (produced) by the above method. The plant may have essentially all of the physiological and morphological characteristics of the deposited line. The above-mentioned "essentially all physiological and morphological characteristics" can be used in the description of the progeny line by replacing "progeny line" with "regenerated plant." Reference 5: Habtamu Gudisa Megersa, “Propagation Methods of Selected Horticultural Crops by Specialized Organs: Review”, Journal of Horticulture, 2017, Volume 4, Issue 2, 1000198

[0252] <Harvested and processed products of Rudbeckia plants> The present disclosure provides harvested and / or processed products of the deposited line or progeny line, wherein the harvested product is a whole plant or part of an individual plant, preferably including flowers; flowers, leaves and / or stems; or seeds.

[0253] The processed product includes any product obtained by processing the deposited line or its progeny line. The processing is not particularly limited, and examples thereof include cutting, slicing, grinding, pureeing, drying, canning, bottling, washing, packaging, freezing, and / or heat treatment. For the deposited line or its progeny line, the plant or plant part used for the processed product may be, for example, a flower. The processed product may be, for example, the deposited line or its progeny line that has been washed and packaged.

[0254] <Method of determining genotype> The present disclosure provides a method for determining or detecting the genotype of a deposited line or a progeny line. The genotype determination method of the present disclosure includes, for example, (a) obtaining a nucleic acid sample from the deposited line or a progeny line, and (b) detecting a genome in the nucleic acid sample. In step (a), nucleic acid preparation from the deposited line or progeny line can be performed using a general nucleic acid preparation method for preparing nucleic acid from tissue. In step (b), for example, polymorphisms and / or alleles in the genome in the nucleic acid sample are detected. The polymorphisms and / or alleles can be detected using, for example, single nucleotide polymorphism (SNP) genotyping, amplified fragment length polymorphism detection (AFLP), restriction fragment length polymorphism discrimination (RFLP) of genomic DNA, sequence-characterized amplified region detection (SCAR) of genomic DNA, cleaved amplified polymorphic sequence detection (CAPS) of genomic DNA, random amplified polymorphic detection (RAPD) of genomic DNA, polymerase chain reaction (PCR), DNA sequencing, allele-specific oligonucleotide (ASO) probes, or DNA microarrays. The polymorphisms and / or alleles can be detected, for example, by sequencing the nucleotide sequence of the genome, or, as described above, by referring to the SNPs of the deposited strain. In step (b), one polymorphism and / or allele in the genomic DNA may be detected, or two or more polymorphisms and / or alleles may be detected. The genotyping method of the present disclosure may include storing the detection results of polymorphisms and / or alleles in a computer-readable medium. The present disclosure may also provide a computer-readable medium produced by such a method. The genome may be, for example, a mitochondrial genome. The genomic DNA may be, for example, mitochondrial genomic DNA.

[0255] The genotyping method of the present disclosure may be performed on any Rudbeckia plant (target Rudbeckia plant) instead of the deposited line or the progeny line. In this case, the genotyping method of the present disclosure may further include, for example, a step of determining whether the target Rudbeckia plant is the progeny line based on the results of step (b). This determination can also be referred to as, for example, discrimination, estimation, appraisal, or identification. The determination can be made, for example, based on the degree of agreement between the results of step (b) and the genotype of the deposited line. [Example]

[0256] The present disclosure will be described in detail below using examples, but the present disclosure is not limited to the aspects described in the examples.

[0257] [Example 1] It was confirmed that a novel sterile Rudbeckia plant exhibits male sterility. Furthermore, a Rudbeckia plant of a deposited line was bred from the male sterile Rudbeckia plant, and the gene responsible for cytoplasmic sterility (SEQ ID NO: 1) was identified from the mitochondrial gene cluster of the deposited line.

[0258] (1) Development of the deposited line and confirmation of cytoplasmic male sterility To develop a novel male-sterile Rudbeckia plant, seeds from a large group of Rudbeckia (Rudbeckia hirta) plants harvested through successive breeding at the Takii Seed Co., Ltd. farm in Konan City, Shiga Prefecture, were bred and tested for sterility. As a result, a novel male-sterile Rudbeckia line (male sterile line) was obtained, which produced no pollen and was presumed to be male sterile. Seeds were obtained by crossing the male-sterile line with pollen from a fertile Rudbeckia plant (Rudbeckia hirta cv. Rowland, fertile line) owned by Takii Seed Co., Ltd. The resulting seeds were bred, and 50 Rudbeckia plants were tested for male sterility using pollen production as an indicator. As a result, all individuals were found to produce no pollen and exhibit male sterility. From the above, it was presumed that the male sterility was cytoplasmic male sterility of maternal origin.

[0259] Next, the sterile line was backcrossed three times using the fertile line as the revertant parent to produce a deposited line (RdCMS line). The deposited line was deposited under accession number FERM BP-22428. In the backcross, all generations of Rudbeckia plants retained male sterility. From these findings, it was determined that the male sterility was maternally inherited cytoplasmic male sterility. Photographs of flowers of the RdCMS line and the fertile line (RdMF) are shown in Figure 1.

[0260] Figure 1 is a photograph showing the flowers of a Rudbeckia plant. In Figure 1, (A) is a photograph of the flowers of the Rudbeckia plant of the deposited line (RdCMS line), and (B) is a photograph of the flowers of the Rudbeckia plant of the fertile line (R. hiruta, RdMF line). As shown in Figure 1, it was found that the fertile line produced pollen, whereas the deposited line did not produce pollen.

[0261] (2) Breeding of cytoplasmic male sterile lines using the deposited lines To breed the cytoplasmic male-sterile lines, fertile lines from multiple Rudbeckia plant groups and the deposited line (RdCMS line) were used. Specifically, the male-fertile lines from the Rudbeckia plant groups were selected from a genetically diverse population. Approximately 5 to 20 individuals sharing the desired agronomic traits were randomly intercrossed, and seeds were collected. Similar selection and crossing were repeated for several generations to obtain fixed male-fertile lines (26 lines). These 26 lines were crossed with the deposited line (RdCMS line) to obtain the F1 generation. For each combination of the F1 generation, 24 individuals were cultivated. As a result, it was found that all individuals from the combination exhibited male sterility (BC1 line). These findings demonstrate that the male sterility gene of the RdCMS line can confer male sterility when combined with various lines. From the 26 lines, a line with high seed production ability (S1 line) was selected. Using the S1 line as a recurrent parent, backcrossing was performed four times with the male sterile line to obtain a backcross progeny line (BC5 line). It was confirmed that the traits of the BC5 line and the S1 line were similar, except for male sterility, and the BC5 line was designated as a male sterile line with fixed traits (SMS1 line). The S1 line was also used as the maintenance line (pollen parent) of the SMS1 line.

[0262] (3) Development of F1 varieties The SMS1 line produced by the method of Example 1(2) was used as the female parent, and a fixed fertile line (SSS line) was used as the male parent, and artificial crossing was performed to obtain a first generation hybrid (F1_MS1 line). The SSS line was a different line from the S1 line. 100 individuals of the F1_MS1 line were cultivated. As a result, all of the individuals exhibited male sterility. A photograph of the F1_MS1 line is shown in Figure 2.

[0263] Figure 2 is a photograph of the F1_MS1 line Rudbeckia plant. As shown in Figure 2, the traits of each individual were uniform, indicating that there was no contamination of self-fertilized seeds. Based on these findings, it was found that cytoplasmic male sterile F1 varieties can be easily bred by using the SMS1 line as a cross parent.

[0264] Furthermore, to establish a more efficient method for producing F1 lineage seeds, we investigated whether honeybees could be used to cross the SMS1 line with the SSS lineage. The SMS1 lineage and the SSS lineage were crossed using honeybees in the same greenhouse where the SMS1 and SSS lines were grown. After crossing using honeybees, seeds were collected from only the female lineage (SMS1 lineage) to obtain the first generation hybrid (F1_MSH lineage). The F1_MS1 lineage and the F1_MSH lineage were sown under the same conditions, and 100 individuals of each were cultivated. No differences were observed between the F1_MSH lineage and the F1_MS1 lineage. These findings demonstrate that it is possible to produce F1 lineage seeds using honeybees.

[0265] (4) Estimation of mitochondrial genes of the deposited strain Cytoplasmic male sterility is known to be caused by mitochondrial genes in the female parent. Therefore, the entire mitochondrial nucleotide sequences of the deposited line (RdCMS line) produced by the method in Example 1(1) and the fertile lines (RdMF line, 26 lines) were identified and compared to infer the causative gene for cytoplasmic male sterility. Specifically, mitochondria were purified from 5 g of fresh leaves of the RdCMS line and the RdMF line. The mitochondrial purification was performed according to the method described in Reference 6 below. After the purification, DNA was extracted from the purified mitochondria. The DNA was extracted according to the method described in Reference 6 below. The extracted DNA from each line was sequenced using the obtained mitochondrial DNA and 10 kb Template Preparation and Sequencing with Low-Input DNA (Pacificaioscience) according to the attached protocol. Specifically, the genomic DNA solution (1 μg equivalent, 150 μl volume) containing the extracted DNA was centrifuged at 6110 rpm for 1 minute using a g-TUBE (Covaris). After centrifugation, the DNA was purified using 0.45 volumes of AMPure PB (Pacificaioscience). After purification, damage in the DNA was repaired, the DNA ends were blunted, and SMRTbell adapters were added. The resulting DNA was then sequenced using the Sequel system (Pacificaioscience). The sequence data obtained from the analysis was then assembled using HGAP4 (Pacificaioscience) and Organella_PBA (see Reference 7 below). The sequence assembly using HGAP4 constructed three virtual genomes for both strains. Furthermore, the sequence assembly using Organella_PBA constructed one virtual genome for both strains. After the assembly, the obtained nucleotide sequence was compared with the nucleotide sequence of the sunflower mitochondrial genome (accession numbers: CM007908 and NC_023337). The sunflower belongs to the Asteraceae family, the same family as the Rudbeckia plant.The base sequences that showed homology with the sunflower mitochondrial genome base sequence were considered to be the base sequences of the hypothetical mitochondrial genomes of each Rudbeckia line, and hypothetical mitochondrial genomes were constructed. The hypothetical mitochondrial genomes constructed for each line were compared. The results are shown in Figure 3. Reference 6: Triboush, et al., “A method for isolation of chloroplast DNA and mitochondrial DNA from sunflower”, Plant molecular biology reporter, 16.2 (1998): 183. Reference 7: Soorni, et al. “Organelle_PBA, a pipeline for assembling chloroplast and mitochondrial genomes from Pacaio DNA sequencing data.”, BMC genomics (2017): 18: 49

[0266] Figure 3 is a graph comparing the mitochondrial genomes of each strain constructed using the two programs. In Figure 3, (A) shows the results for the hypothetical mitochondrial genome of the deposited strain (RdCMS strain), and (B) shows the results for the hypothetical mitochondrial genome of the normal cytoplasmic strain (RdMF strain). In Figures 3(A) and (B), the vertical axis shows the hypothetical mitochondrial genome constructed using Organella_PBA, and the horizontal axis shows the hypothetical mitochondrial genome constructed using HGAP4. As shown in Figure 3, the hypothetical mitochondrial genomes constructed using Organella_PBA and HGAP4 for both the RdCMS strain and the RdMF strain were nearly identical.

[0267] It has been suggested that cytoplasmic male sterility genes arise from chimeric genes or loss-of-function mutations (Reference 8). Therefore, to identify genetic variations between the RdCMS and RdMF lines, gene prediction was performed for each hypothetical mitochondrial genome based on information on known mitochondrial genes. Mitofy (Reference 9) was used for gene prediction. Furthermore, gene prediction was performed for each hypothetical mitochondrial genome using getORF (https: / / www.bioinformatics.nl / cgi-bin / emboss / getorf). For genes predicted from each hypothetical genome obtained from Mitofy and getORF, CD-HIT (https: / / github.com / weizhongli / cdhit) was used to construct unique gene sets based on the amino acid sequences. Specifically, genes showing 90% or more homology over 50% or more of the amino acid sequence of each gene were considered to be overlapping identical genes. As a result, a total of 47 genes were predicted from the RdCMS and RdMF strains. The locations of the 47 genes on each hypothetical mitochondrial genome were visualized using Simple Synteny (Reference 10). Reference 8: Touzet, Pascal, and Etienne H. Meyer. "Cytoplasmic male sterility and mitochondrial metabolism in plants." Mitochondrion 19 (2014): 166-171. Reference 9: Alverson, et al., "Insights into the evolution of mitochondrial genome size from complete sequences of Citrullus lanatus and Cucurbita pepo (Cucurbitaceae)." Molecular biology and evolution 27.6 (2010): 1436-1448. Reference 10: Veltri, et al., "SimpleSynteny: a web-based tool for visualization of microsynteny across multiple species." Nucleicacids research 44.W1 (2016): W41-W45.

[0268] Next, the amino acid sequences of the 47 genes predicted from the RdCMS and RdMF strains were compared using BLAST (https: / / blast.ncai.nlm.nih.gov / Blast.cgi). Specifically, a database was created for the hypothetical mitochondrial genome of each strain. Homology analysis was performed using the database as the subject and the 47 genes predicted from the RdCMS and RdMF strains as queries. Genes that showed 90% or more homology over 60% or more of the amino acid sequence length of each gene between the two strains were defined as genes commonly present in both strains. Genes other than the commonly present genes were defined as genes specific to one of the two strains. These results are shown in Figure 4.

[0269] Figure 4 shows the genes predicted on the mitochondrial genome of each strain, constructed using Organella_PBA or HGAP4. As shown in Figure 4, of the 47 gene sets, 40 gene sets were genes commonly present in both strains. Furthermore, of the 47 gene sets, 7 gene sets were genes not commonly present in both strains. For some genes, the gene arrangement was identical between the RdCMS strain and the RdMF strain. For many genes, the gene arrangement between the RdCMS strain and the RdMF strain was not conserved.

[0270] Furthermore, of the seven genes that were present only in one of the strains, six genes (ORF1, ORF2, ORF3, ORF6, ORF7, and RPS7) were present in the RdCMS strain but not in the RdMF strain. ORF1 showed high homology to DNA-dependent RNA polymerase. ORF2 and ORF6 showed high homology to DNA polymerase. ORF3 was a gene showing homology to ATP6, which has an ATP synthesis domain. ORF7 was a gene of unknown function. Of the seven genes that were present only in one of the strains, one gene (ATP6) was present in the RdMF strain but not in the RdCMS strain.

[0271] ATP6 is known to be the cause of cytoplasmic male sterility (CMS) in some plants. Furthermore, among the genes specifically present in the RdCMS line, there was a gene ORF3 that showed homology to the gene (ATP6) specifically present in the RdMF line. Therefore, it was thought that ORF3, which has homology to ATP6, contributes to cytoplasmic male sterility (CMS). Therefore, the ATP6 (SEQ ID NO: 3) found in the RdMF line and the ORF3 (SEQ ID NO: 1) found in the RdCMS line were compared. corresponds to the latter half of ORF3 in Figure 5 (the base sequence up to the translation stop codon) The results of the comparison are shown in Figure 5.

[0272] Figure 5 is a schematic diagram comparing the nucleotide sequences of ATP6 from the RdMF strain and ORF3 from the RdCMS strain. As shown in Figure 5, when the translation initiation site is defined as the first amino acid, the nucleotide sequence from positions 763 to 1512 of the translation stop codon in ATP6 is completely identical to the nucleotide sequence from positions 340 to 1089 of the translation stop codon in ORF3. Furthermore, the nucleotide sequence from positions 1 to 762 of ATP6 is not identical to the nucleotide sequence from positions 1 to 339 of ORF3. The nucleotide sequence from positions 1 to 339 of ORF3, which did not show any identity with ATP6, and the nucleotide sequence from positions 1 to 679 upstream of the translation initiation site (position 0) (SEQ ID NO: 8), were identical to the nucleotide sequence from positions 24133 to 24819 of the hypothetical mitochondrial genome of the RdMF strain constructed with HGAP4 (contig: MF1_00001F, SEQ ID NO: 9). These findings suggest that ORF3 in the RdCMS strain is a chimeric gene in which ATP6 is fused with the base sequence of another region present in the RdMF strain.

[0273] The base sequence around the translation start point of ORF3 (base sequence of SEQ ID NO: 18)

[0274] (5) Identification of the gene responsible for cytoplasmic sterility in the deposited line We investigated whether the male sterility of the deposited line (RdCMS line) was due to differences in ORF3 and ATP6. Specifically, PCR was used to examine whether ORF3 or ATP6 was detected in the RdCMS line and other male-fertile Rudbeckia plants. First, DNA was extracted from approximately 50 mg of leaves from the RdCMS line, the RdMF line, male-fertile Rudbeckia plant lines (120 lines) owned by Takii Seed Co., Ltd., and male-fertile Rudbeckia plants (48 individuals) growing wild within Takii Seed Co., Ltd. DNA was extracted using a DNA extraction kit (Puregene DNA, manufactured by QIAGEN) according to the attached protocol. A reaction mixture containing 1 μg of the resulting DNA, 0.5 μl of 10× buffer, 0.25 μl of HsExTaq (Takara Bio), 0.5 μl of primer mixture, and 2.75 μl of purified water was used to amplify DNA using a PCR system (Takara). The primer mixture was prepared so that each primer in the ORF3 primer set or ATP6 primer set below was 20 mmol / L. The PCR conditions were 95°C for 10 minutes, followed by 35 cycles of 95°C for 1 minute, 58°C for 30 seconds, and 72°C for 30 seconds. After DNA amplification, the reaction mixture was electrophoresed on a 1% agarose gel to confirm the amplified DNA. Representative examples of these results are shown in Figure 6.

[0275] Primer set for ORF3 Forward primer (SEQ ID NO: 14) 5'- ACGTAACACGTATCTATGGTGCAT-3' Reverse primer (SEQ ID NO: 15) 5'-GAGAGTTAGCAGCATAAACAAAGA-3' Primer set for ATP6 Forward primer (SEQ ID NO: 16) 5'- GAGGGACTTTATTCAGTCTTATCG-3' Reverse primer (SEQ ID NO: 17) 5'- ATCTTCATAGGAATCAATGGGAGA-3'

[0276] Figure 6 shows photographs of the electrophoresis results for fertile lines (R. hiruta, RdMF line), deposited lines (RdCMS line), R. hiruta (120 lines) owned by Takii Seed Co., Ltd., and R. hiruta plants growing wild within Takii Seed Co., Ltd. In Figure 6, the samples, from left to right, are the RdMF line (M), the RdCMS line (C), and commercially available fertile lines (1-8). The upper row in Figure 6 shows the results for ORF3, and the lower row shows the results for ATP6. All lines except the deposited line (RdCMS line) are fertile. As shown in Figure 6, DNA amplification was observed in all RdCMs line individuals when primers for ORF3 were used, whereas no DNA amplification was observed when primers for ATP6 were used. On the other hand, in all 120 fertile lines and 48 wild individuals, no DNA amplification was observed when ORF3 primers were used, whereas DNA amplification was observed when ATP6 primers were used. Similar results were also obtained in fertile lines not shown in Figure 6. From these results, it was found that the presence or absence of ATP6 and ORF3 correlates with the presence or absence of male sterility.

[0277] These results suggest that ORF3 of the deposited line (RdCMS line) is the gene responsible for cytoplasmic sterility in Rudbeckia plants. Furthermore, it was shown that cytoplasmic sterility derived from the RdCMS line can be efficiently selected using the ATP6 primer and ORF3 primer used in Example 1(5).

[0278] Although the present disclosure has been described above with reference to the embodiments and examples, the present disclosure is not limited to the above-described embodiments and examples. Various modifications that can be understood by those skilled in the art can be made to the configuration and details of the present disclosure within the scope of the present disclosure.

[0279] <Additional Notes> Some or all of the above-described embodiments and examples can be described as, but are not limited to, the following supplementary notes. <Male sterile Rudbeckia plant> (Appendix 1) Rudbeckia plants that are cytoplasmic male sterile. (Appendix 2) A Rudbeckia plant according to Appendix 1, comprising, as a cytoplasmic male sterility gene, at least one polynucleotide selected from the group consisting of the following (ca) to (ce) and (cf): (ca) any one of the following polynucleotides (ca1) to (ca7): (ca1) a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1; (ca2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (ca1); (ca3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (ca1), and that causes cytoplasmic male sterility; (ca4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (ca1); (ca5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 2; (ca6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 2 in which one or several amino acids have been deleted, substituted, inserted and / or added; (ca7) a polynucleotide that expresses cytoplasmic male sterility and that consists of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 2; (cb) any one of the following polynucleotides (cb1) to (cb7): (cb1) a polynucleotide consisting of the base sequence of SEQ ID NO: 3; (cb2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cb1); (cb3) a polynucleotide that expresses cytoplasmic male sterility and that consists of a base sequence having 80% or more identity to the base sequence of (cb1); (cb4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cb1); (cb5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 4; (cb6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 4 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cb7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 4; (cc) any one of the following polynucleotides (cc1) to (cc7): (cc1) a polynucleotide consisting of the base sequence of SEQ ID NO: 5; (cc2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cc1); (cc3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (cc1), and that expresses cytoplasmic male sterility; (cc4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to the polynucleotide having the nucleotide sequence of (cc1); (cc5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 6; (cc6) A polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 6 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cc7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 6; (cd) any one of the following polynucleotides (cd1) to (cd7): (cd1) a polynucleotide consisting of the base sequence of SEQ ID NO: 7; (cd2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cd1); (cd3) a polynucleotide consisting of a base sequence having 80% or more identity to the base sequence of (cd1), which expresses cytoplasmic male sterility; (cd4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cd1); (cd5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 8; (cd6) A polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 8 in which one or several amino acids have been deleted, substituted, inserted and / or added; (cd7) a polynucleotide that expresses cytoplasmic male sterility and consists of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 8; (ce) any one of the following polynucleotides (ce1) to (ce7): (ce1) a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 9; (ce2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (ce1); (ce3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (ce1), and that causes cytoplasmic male sterility; (ce4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the nucleotide sequence of (ce1); (ce5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 10; (ce6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 10 in which one or several amino acids have been deleted, substituted, inserted and / or added; (ce7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 10; (cf) any one of the following polynucleotides (cf1) to (cf7): (cf1) a polynucleotide consisting of the base sequence of SEQ ID NO: 11; (cf2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cf1); (cf3) a polynucleotide that expresses cytoplasmic male sterility and that consists of a base sequence having 80% or more identity to the base sequence of (cf1); (cf4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to the polynucleotide having the nucleotide sequence of (cf1); (cf5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 12; (cf6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 12 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cf7) A polynucleotide consisting of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 12, which polynucleotide causes expression of cytoplasmic male sterility. (Appendix 3) 3. The Rudbeckia plant according to Appendix 2, comprising the polynucleotide (ca) as the cytoplasmic male sterility gene. (Appendix 4) 4. The Rudbeckia plant according to claim 2 or 3, comprising the polynucleotides (ca) to (ce) and (cf) as the cytoplasmic male sterility gene. (Appendix 5) 5. A Rudbeckia plant according to any one of Appendices 2 to 4, comprising the cytoplasmic male sterility gene in its mitochondrial genome. (Appendix 6) 6. The Rudbeckia plant of any one of Appendices 1 to 5, wherein the Rudbeckia plant is a plant grown from seeds deposited under Accession No. FERM BP-22428. (Appendix 7) A progeny line of the Rudbeckia plant described in any one of Supplementary Notes 1 to 6, The progeny line exhibits cytoplasmic male sterility. (Appendix 8) 8. The progeny line of claim 7, wherein the progeny line is a first generation hybrid line. (Appendix 9) 10. Seeds of a Rudbeckia plant according to any one of appendices 1 to 6, or a progeny line according to appendices 7 or 8. (Appendix 10) 10. A part of a Rudbeckia plant according to any one of appendices 1 to 6, or a progeny line according to appendices 7 or 8. (Appendix 11) 10. A callus comprising cells of a Rudbeckia plant according to any one of appendices 1 to 6, or a progeny line according to appendices 7 or 8. (Appendix 12) Cytoplasm contained in a Rudbeckia plant according to any one of Appendixes 1 to 6, a progeny line according to Appendix 7 or 8, a seed according to Appendix 9, a part according to Appendix 10, or a callus according to Appendix 11. <Method for producing male sterile Rudbeckia plants (first production method)> (Appendix 13) A method for producing a cytoplasmically male sterile Rudbeckia plant, comprising the following step (a): (a) crossing a Rudbeckia plant according to any one of Appendixes 1 to 6, or a progeny line according to Appendix 7 or 8, with another Rudbeckia plant. (Appendix 14) The production method according to Appendix 13, further comprising the following step (x) prior to the step (a): (x) selecting a Rudbeckia plant according to any one of Appendices 1 to 6 or a progeny line according to Appendices 7 or 8 from a test Rudbeckia plant. (Appendix 15) The method for producing Rudbeckia plants according to Appendix 14, wherein in the step (x), the Rudbeckia plants are selected which contain, as a cytoplasmic male sterility gene, at least one polynucleotide selected from the group consisting of the following (ca) to (ce) and (cf): (ca) any one of the following polynucleotides (ca1) to (ca7): (ca1) a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1; (ca2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (ca1); (ca3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (ca1), and that causes cytoplasmic male sterility; (ca4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (ca1); (ca5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 2; (ca6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 2 in which one or several amino acids have been deleted, substituted, inserted and / or added; (ca7) a polynucleotide that expresses cytoplasmic male sterility and that consists of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 2; (cb) any one of the following polynucleotides (cb1) to (cb7): (cb1) a polynucleotide consisting of the base sequence of SEQ ID NO: 3; (cb2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cb1); (cb3) a polynucleotide that expresses cytoplasmic male sterility and that consists of a base sequence having 80% or more identity to the base sequence of (cb1); (cb4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cb1); (cb5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 4; (cb6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 4 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cb7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 4; (cc) any one of the following polynucleotides (cc1) to (cc7): (cc1) a polynucleotide consisting of the base sequence of SEQ ID NO: 5; (cc2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cc1); (cc3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (cc1), and that expresses cytoplasmic male sterility; (cc4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to the polynucleotide having the nucleotide sequence of (cc1); (cc5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 6; (cc6) A polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 6 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cc7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 6; (cd) any one of the following polynucleotides (cd1) to (cd7): (cd1) a polynucleotide consisting of the base sequence of SEQ ID NO: 7; (cd2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cd1); (cd3) a polynucleotide consisting of a base sequence having 80% or more identity to the base sequence of (cd1), which expresses cytoplasmic male sterility; (cd4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cd1); (cd5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 8; (cd6) A polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 8 in which one or several amino acids have been deleted, substituted, inserted and / or added; (cd7) a polynucleotide that expresses cytoplasmic male sterility and consists of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 8; (ce) any one of the following polynucleotides (ce1) to (ce7): (ce1) a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 9; (ce2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (ce1); (ce3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (ce1), and that causes cytoplasmic male sterility; (ce4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the nucleotide sequence of (ce1); (ce5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 10; (ce6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 10 in which one or several amino acids have been deleted, substituted, inserted and / or added; (ce7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 10; (cf) any one of the following polynucleotides (cf1) to (cf7): (cf1) a polynucleotide consisting of the base sequence of SEQ ID NO: 11; (cf2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cf1); (cf3) a polynucleotide that expresses cytoplasmic male sterility and that consists of a base sequence having 80% or more identity to the base sequence of (cf1); (cf4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to the polynucleotide having the nucleotide sequence of (cf1); (cf5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 12; (cf6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 12 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cf7) A polynucleotide consisting of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 12, which polynucleotide causes expression of cytoplasmic male sterility. (Appendix 16) 16. The method according to claim 15, wherein in the step (x), a Rudbeckia plant containing the polynucleotide of (ca) as the cytoplasmic male sterility gene is selected. (Appendix 17) 17. The method according to claim 15 or 16, wherein in the step (x), a Rudbeckia plant containing any one of the polynucleotides (ca) to (cf) as the cytoplasmic male sterility gene is selected. (Appendix 18) 18. The production method according to any one of Appendices 15 to 17, wherein in step (x), the cytoplasmic male sterility gene is detected in the test Rudbeckia plant, and a test Rudbeckia plant containing the cytoplasmic male sterility gene is selected. (Appendix 19) 19. The production method described in any one of Appendices 15 to 18, wherein in step (x), the cytoplasmic male sterility gene is detected in the mitochondrial genome of the test Rudbeckia plant, and a test Rudbeckia plant containing the cytoplasmic male sterility gene is selected. (Appendix 20) A production method according to any one of Appendices 13 to 19, comprising the following step (b): (b) A step of selecting a cytoplasmic male sterile Rudbeckia plant from the Rudbeckia plant obtained in the step (a) or its progeny line. (Appendix 21) The method according to Appendix 20, wherein in the step (b), a Rudbeckia plant is selected which contains, as a cytoplasmic male sterility gene, at least one polynucleotide selected from the group consisting of the following (ca) to (ce) and (cf): (ca) any one of the following polynucleotides (ca1) to (ca7): (ca1) a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1; (ca2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (ca1); (ca3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (ca1), and that causes cytoplasmic male sterility; (ca4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (ca1); (ca5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 2; (ca6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 2 in which one or several amino acids have been deleted, substituted, inserted and / or added; (ca7) a polynucleotide that expresses cytoplasmic male sterility and that consists of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 2; (cb) any one of the following polynucleotides (cb1) to (cb7): (cb1) a polynucleotide consisting of the base sequence of SEQ ID NO: 3; (cb2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cb1); (cb3) a polynucleotide that expresses cytoplasmic male sterility and that consists of a base sequence having 80% or more identity to the base sequence of (cb1); (cb4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cb1); (cb5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 4; (cb6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 4 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cb7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 4; (cc) any one of the following polynucleotides (cc1) to (cc7): (cc1) a polynucleotide consisting of the base sequence of SEQ ID NO: 5; (cc2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cc1); (cc3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (cc1), and that expresses cytoplasmic male sterility; (cc4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to the polynucleotide having the nucleotide sequence of (cc1); (cc5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 6; (cc6) A polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 6 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cc7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 6; (cd) any one of the following polynucleotides (cd1) to (cd7): (cd1) a polynucleotide consisting of the base sequence of SEQ ID NO: 7; (cd2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cd1); (cd3) a polynucleotide consisting of a base sequence having 80% or more identity to the base sequence of (cd1), which expresses cytoplasmic male sterility; (cd4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cd1); (cd5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 8; (cd6) A polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 8 in which one or several amino acids have been deleted, substituted, inserted and / or added; (cd7) a polynucleotide that expresses cytoplasmic male sterility and consists of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 8; (ce) any one of the following polynucleotides (ce1) to (ce7): (ce1) a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 9; (ce2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (ce1); (ce3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (ce1), and that causes cytoplasmic male sterility; (ce4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the nucleotide sequence of (ce1); (ce5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 10; (ce6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 10 in which one or several amino acids have been deleted, substituted, inserted and / or added; (ce7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 10; (cf) any one of the following polynucleotides (cf1) to (cf7): (cf1) a polynucleotide consisting of the base sequence of SEQ ID NO: 11; (cf2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cf1); (cf3) a polynucleotide that expresses cytoplasmic male sterility and that consists of a base sequence having 80% or more identity to the base sequence of (cf1); (cf4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to the polynucleotide having the nucleotide sequence of (cf1); (cf5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 12; (cf6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 12 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cf7) A polynucleotide consisting of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 12, which polynucleotide causes expression of cytoplasmic male sterility. (Appendix 22) 22. The method according to claim 21, wherein in the step (b), a Rudbeckia plant containing the polynucleotide of (ca) as the cytoplasmic male sterility gene is selected. (Appendix 23) 23. The method according to claim 21 or 22, wherein in the step (b), a Rudbeckia plant containing any one of the polynucleotides (ca) to (cf) as the cytoplasmic male sterility gene is selected. (Appendix 24) 24. The production method according to any one of Appendices 21 to 23, wherein in step (b), the cytoplasmic male sterility gene is detected in the test Rudbeckia plant, and a test Rudbeckia plant containing the cytoplasmic male sterility gene is selected. (Appendix 25) A production method described in any of Appendices 21 to 24, wherein in step (b), the cytoplasmic male sterility gene is detected in the mitochondrial genome of the test Rudbeckia plant, and a test Rudbeckia plant containing the cytoplasmic male sterility gene is selected. <Method for imparting male sterility to Rudbeckia plants> (Appendix 26) A method for imparting cytoplasmic male sterility to a Rudbeckia plant, comprising the step of introducing into a target Rudbeckia plant at least one polynucleotide selected from the group consisting of the following (ca) to (ce) and (cf) as a cytoplasmic male sterility gene: (ca) any one of the following polynucleotides (ca1) to (ca7): (ca1) a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1; (ca2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (ca1); (ca3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (ca1), and that causes cytoplasmic male sterility; (ca4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (ca1); (ca5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 2; (ca6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 2 in which one or several amino acids have been deleted, substituted, inserted and / or added; (ca7) a polynucleotide that expresses cytoplasmic male sterility and that consists of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 2; (cb) any one of the following polynucleotides (cb1) to (cb7): (cb1) a polynucleotide consisting of the base sequence of SEQ ID NO: 3; (cb2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cb1); (cb3) a polynucleotide that expresses cytoplasmic male sterility and that consists of a base sequence having 80% or more identity to the base sequence of (cb1); (cb4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cb1); (cb5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 4; (cb6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 4 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cb7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 4; (cc) any one of the following polynucleotides (cc1) to (cc7): (cc1) a polynucleotide consisting of the base sequence of SEQ ID NO: 5; (cc2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cc1); (cc3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (cc1), and that expresses cytoplasmic male sterility; (cc4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to the polynucleotide having the nucleotide sequence of (cc1); (cc5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 6; (cc6) A polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 6 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cc7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 6; (cd) any one of the following polynucleotides (cd1) to (cd7): (cd1) a polynucleotide consisting of the base sequence of SEQ ID NO: 7; (cd2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cd1); (cd3) a polynucleotide consisting of a base sequence having 80% or more identity to the base sequence of (cd1), which expresses cytoplasmic male sterility; (cd4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cd1); (cd5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 8; (cd6) A polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 8 in which one or several amino acids have been deleted, substituted, inserted and / or added; (cd7) a polynucleotide that expresses cytoplasmic male sterility and consists of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 8; (ce) any one of the following polynucleotides (ce1) to (ce7): (ce1) a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 9; (ce2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (ce1); (ce3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (ce1), and that causes cytoplasmic male sterility; (ce4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the nucleotide sequence of (ce1); (ce5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 10; (ce6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 10 in which one or several amino acids have been deleted, substituted, inserted and / or added; (ce7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 10; (cf) any one of the following polynucleotides (cf1) to (cf7): (cf1) a polynucleotide consisting of the base sequence of SEQ ID NO: 11; (cf2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cf1); (cf3) a polynucleotide that expresses cytoplasmic male sterility and that consists of a base sequence having 80% or more identity to the base sequence of (cf1); (cf4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to the polynucleotide having the nucleotide sequence of (cf1); (cf5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 12; (cf6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 12 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cf7) A polynucleotide consisting of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 12, which polynucleotide causes expression of cytoplasmic male sterility. (Appendix 27) 27. The method of imparting cytoplasmic male sterility according to claim 26, wherein the introducing step involves introducing a Rudbeckia plant containing the polynucleotide (ca) as the cytoplasmic male sterility gene. (Appendix 28) 28. The method for imparting cytoplasmic male sterility according to claim 26 or 27, wherein the introduction step involves introducing a Rudbeckia plant containing any one of the polynucleotides (ca) to (cf) as the cytoplasmic male sterility gene. (Appendix 29) 29. The method of any one of Appendices 26 to 28, wherein in the introducing step, the cytoplasmic male sterility gene is introduced into the mitochondrial genome of the target Rudbeckia plant. <Method for producing male sterile Rudbeckia plants (second production method)> (Appendix 30) The method includes a step of conferring cytoplasmic male sterility to a target Rudbeckia plant, A method for producing a cytoplasmically male sterile Rudbeckia plant, wherein the conferring step is carried out by the conferring method described in any one of Appendices 26 to 29. <Method for screening male sterile Rudbeckia plants> (Appendix 31) A method for screening for cytoplasmic male sterile Rudbeckia plants, the method comprising a selection step of selecting, from test Rudbeckia plants, test Rudbeckia plants that contain, as a cytoplasmic male sterility gene, at least one polynucleotide selected from the group consisting of the following (ca) to (ce) and (cf): (ca) any one of the following polynucleotides (ca1) to (ca7): (ca1) a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1; (ca2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (ca1); (ca3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (ca1), and that causes cytoplasmic male sterility; (ca4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (ca1); (ca5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 2; (ca6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 2 in which one or several amino acids have been deleted, substituted, inserted and / or added; (ca7) a polynucleotide that expresses cytoplasmic male sterility and that consists of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 2; (cb) any one of the following polynucleotides (cb1) to (cb7): (cb1) a polynucleotide consisting of the base sequence of SEQ ID NO: 3; (cb2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cb1); (cb3) a polynucleotide that expresses cytoplasmic male sterility and that consists of a base sequence having 80% or more identity to the base sequence of (cb1); (cb4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cb1); (cb5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 4; (cb6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 4 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cb7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 4; (cc) any one of the following polynucleotides (cc1) to (cc7): (cc1) a polynucleotide consisting of the base sequence of SEQ ID NO: 5; (cc2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cc1); (cc3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (cc1), and that expresses cytoplasmic male sterility; (cc4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to the polynucleotide having the nucleotide sequence of (cc1); (cc5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 6; (cc6) A polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 6 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cc7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 6; (cd) any one of the following polynucleotides (cd1) to (cd7): (cd1) a polynucleotide consisting of the base sequence of SEQ ID NO: 7; (cd2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cd1); (cd3) a polynucleotide consisting of a base sequence having 80% or more identity to the base sequence of (cd1), which expresses cytoplasmic male sterility; (cd4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cd1); (cd5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 8; (cd6) A polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 8 in which one or several amino acids have been deleted, substituted, inserted and / or added; (cd7) a polynucleotide that expresses cytoplasmic male sterility and consists of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 8; (ce) any one of the following polynucleotides (ce1) to (ce7): (ce1) a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 9; (ce2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (ce1); (ce3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (ce1), and that causes cytoplasmic male sterility; (ce4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the nucleotide sequence of (ce1); (ce5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 10; (ce6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 10 in which one or several amino acids have been deleted, substituted, inserted and / or added; (ce7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 10; (cf) any one of the following polynucleotides (cf1) to (cf7): (cf1) a polynucleotide consisting of the base sequence of SEQ ID NO: 11; (cf2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cf1); (cf3) a polynucleotide that expresses cytoplasmic male sterility and that consists of a base sequence having 80% or more identity to the base sequence of (cf1); (cf4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to the polynucleotide having the nucleotide sequence of (cf1); (cf5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 12; (cf6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 12 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cf7) A polynucleotide consisting of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 12, which polynucleotide causes expression of cytoplasmic male sterility. (Appendix 32) 32. The screening method according to claim 31, wherein the selection step comprises selecting a Rudbeckia plant comprising the polynucleotide (ca) as the cytoplasmic male sterility gene. (Appendix 33) 33. The screening method according to claim 31 or 32, wherein the selection step involves selecting a Rudbeckia plant containing any one of the polynucleotides (ca) to (cf) as the cytoplasmic male sterility gene. (Appendix 34) A screening method described in any of Appendices 31 to 33, wherein the selection step involves detecting the cytoplasmic male sterility gene in the test Rudbeckia plant and selecting a test Rudbeckia plant containing the cytoplasmic male sterility gene. (Appendix 35) A screening method described in any of Appendices 31 to 34, wherein the selection step involves detecting the cytoplasmic male sterility gene in the mitochondrial genome of the test Rudbeckia plant and selecting a test Rudbeckia plant containing the cytoplasmic male sterility gene. <Method for producing male sterile Rudbeckia plants (third production method)> (Appendix 36) a screening step of screening test Rudbeckia plants containing a cytoplasmic male sterility gene from test Rudbeckia plants; A method for producing a cytoplasmically male sterile Rudbeckia plant, wherein the screening step is carried out by the screening method described in any one of Appendices 31 to 35. <Male sterile Rudbeckia plant> (Appendix 37) A cytoplasmically male sterile Rudbeckia plant obtainable by the production method of any of Appendixes 13 to 25, the production method of Appendix 30, or the production method of Appendix 36. <Method for detecting male sterility in Rudbeckia plants> (Appendix 38) A method for detecting cytoplasmic male sterility in a Rudbeckia plant, comprising a detection step of detecting, in a test Rudbeckia plant, at least one polynucleotide selected from the group consisting of the following (ca) to (ce) and (cf) as a cytoplasmic male sterility gene: (ca) any one of the following polynucleotides (ca1) to (ca7): (ca1) a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1; (ca2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (ca1); (ca3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (ca1), and that causes cytoplasmic male sterility; (ca4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (ca1); (ca5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 2; (ca6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 2 in which one or several amino acids have been deleted, substituted, inserted and / or added; (ca7) a polynucleotide that expresses cytoplasmic male sterility and that consists of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 2; (cb) any one of the following polynucleotides (cb1) to (cb7): (cb1) a polynucleotide consisting of the base sequence of SEQ ID NO: 3; (cb2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cb1); (cb3) a polynucleotide that expresses cytoplasmic male sterility and that consists of a base sequence having 80% or more identity to the base sequence of (cb1); (cb4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cb1); (cb5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 4; (cb6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 4 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cb7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 4; (cc) any one of the following polynucleotides (cc1) to (cc7): (cc1) a polynucleotide consisting of the base sequence of SEQ ID NO: 5; (cc2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cc1); (cc3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (cc1), and that expresses cytoplasmic male sterility; (cc4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to the polynucleotide having the nucleotide sequence of (cc1); (cc5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 6; (cc6) A polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 6 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cc7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 6; (cd) any one of the following polynucleotides (cd1) to (cd7): (cd1) a polynucleotide consisting of the base sequence of SEQ ID NO: 7; (cd2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cd1); (cd3) a polynucleotide consisting of a base sequence having 80% or more identity to the base sequence of (cd1), which expresses cytoplasmic male sterility; (cd4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cd1); (cd5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 8; (cd6) A polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 8 in which one or several amino acids have been deleted, substituted, inserted and / or added; (cd7) a polynucleotide that expresses cytoplasmic male sterility and consists of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 8; (ce) any one of the following polynucleotides (ce1) to (ce7): (ce1) a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 9; (ce2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (ce1); (ce3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (ce1), and that causes cytoplasmic male sterility; (ce4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the nucleotide sequence of (ce1); (ce5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 10; (ce6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 10 in which one or several amino acids have been deleted, substituted, inserted and / or added; (ce7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 10; (cf) any one of the following polynucleotides (cf1) to (cf7): (cf1) a polynucleotide consisting of the base sequence of SEQ ID NO: 11; (cf2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cf1); (cf3) a polynucleotide that expresses cytoplasmic male sterility and that consists of a base sequence having 80% or more identity to the base sequence of (cf1); (cf4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to the polynucleotide having the nucleotide sequence of (cf1); (cf5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 12; (cf6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 12 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cf7) A polynucleotide consisting of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 12, which polynucleotide causes expression of cytoplasmic male sterility. (Appendix 39) 39. The detection method according to claim 38, wherein the detection step detects the polynucleotide of (ca) as the cytoplasmic male sterility gene. (Appendix 40) 40. The detection method according to claim 38 or 39, wherein the polynucleotides (ca) to (cf) are detected as the cytoplasmic male sterility gene in the selection step. <First deposited strain> (Appendix 41) Seeds of Rudbeckia plants deposited under accession number FERM BP-22428. (Appendix 42) A Rudbeckia plant grown from seeds of the Rudbeckia plant deposited under accession number FERM BP-22428. (Appendix 43) A progeny line of a Rudbeckia plant described in Appendix 41 or 42. (Appendix 44) 44. The progeny line of claim 43, wherein the progeny line is a first generation hybrid line. (Appendix 45) 4. Seeds of a Rudbeckia plant according to claim 41 or 42, or a progeny line according to claim 43 or 44. (Appendix 46) A Rudbeckia plant according to appendix 41 or 42, or a portion of a progeny line according to appendix 43 or 44. (Appendix 47) 46. ​​A callus comprising cells of a Rudbeckia plant according to claim 41 or 42, or a progeny line according to claim 43 or 44. (Appendix 48) 46. ​​Cytoplasm contained in a Rudbeckia plant according to Appendix 41 or 42, a progeny line according to Appendix 43 or 44, a seed according to Appendix 45, a part according to Appendix 46, or a callus according to Appendix 47. (Appendix 49) 46. ​​Mitochondria contained in a Rudbeckia plant according to appendix 41 or 42, a progeny line according to appendix 43 or 44, a seed according to appendix 45, a part according to appendix 46, or a callus according to appendix 47. (Appendix 50) 46. ​​A mitochondrial genome contained in a Rudbeckia plant according to Appendix 41 or 42, a progeny line according to Appendix 43 or 44, a seed according to Appendix 45, a part according to Appendix 46, or a callus according to Appendix 47. (Appendix 51) A method for producing a Rudbeckia plant, comprising a crossbreeding step of crossing a Rudbeckia plant according to Appendix 41 or 42, or a progeny line according to Appendix 43 or 44, with another Rudbeckia plant. (Appendix 52) A method for producing a Rudbeckia plant according to claim 51, comprising a seed collection step of collecting seeds. <Second deposited strain> (Appendix 53) Rudbeckia cultivar Takii 22 seeds, A representative sample is a seed of the Rudbeckia plant deposited under accession number FERM BP-22428. (Appendix 54) Rudbeckia species Takii 22 Rudbeckia plants, Rudbeckia plants, a representative sample of which is the seed of the Rudbeckia plant deposited under accession number FERM BP-22428. (Appendix 55) Rudbeckia plants or parts thereof, 54. A Rudbeckia plant or part thereof, wherein the Rudbeckia plant or part thereof has essentially all of the physiological and morphological characteristics of the Rudbeckia plant described in Appendix 54. (Appendix 56) A progeny Rudbeckia plant of the Rudbeckia plant described in Appendix 54, The progeny Rudbeckia plant contains at least 50% of the alleles of the Rudbeckia plant described in Appendix 54; The progeny Rudbeckia plant is a progeny Rudbeckia plant having cytoplasmic male sterility. (Appendix 57) 56. Seeds that produce the Rudbeckia plants described in Appendix 56. (Appendix 58) Parts of the Rudbeckia plant described in Appendix 54. (Appendix 59) 58. A plant part according to claim 58, The plant part includes an ovary, an ovule, an egg cell, a cutting, a root, a stem, a leaf, a cell or a protoplast. (Appendix 60) A method for producing Rudbeckia seeds, The method comprises: Crossing the Rudbeckia plant described in Appendix 54 with another Rudbeckia plant, harvesting the resulting seeds. (Appendix 61) 61. Rudbeckia seeds derived from a Rudbeckia plant, produced by the method described in Appendix 60. (Appendix 62) 6. A Rudbeckia plant or part thereof produced by growing the Rudbeckia seeds described in Appendix 61. (Appendix 63) 63. A Rudbeckia plant or part thereof according to claim 62, the Rudbeckia plant or part thereof contains at least 50% alleles of Rudbeckia cultivar Takii 22, a representative sample of which is seed of the Rudbeckia plant deposited under accession number FERM BP-22428; The Rudbeckia plant or part thereof is cytoplasmic male sterility. (Appendix 64) 64. A Rudbeckia plant or part thereof according to claim 63, The Rudbeckia plant or part thereof has been modified in one or more traits. (Appendix 65) 65. A Rudbeckia plant or part thereof according to claim 64, A Rudbeckia plant or part thereof, wherein said modification is effected by mutagenesis. (Appendix 66) 55. A method for producing seeds of a Rudbeckia plant from the Rudbeckia plant of claim 54, comprising: The method comprises: (a) crossing Rudbeckia cultivar Takii 22, which is the seed of the Rudbeckia plant deposited under accession number FERM BP-22428, with another Rudbeckia plant to produce seed; (b) growing Rudbeckia plants from the seeds of step (a) to produce Rudbeckia plants derived from the Rudbeckia cultivar Takii 22; (c) crossing the Rudbeckia plant of step (b) with another Rudbeckia plant to produce an additional Rudbeckia plant derived from Rudbeckia cultivar Takii 22; (d) optionally repeating steps (b) and (c) one or more times to produce additional Rudbeckia plants derived from Rudbeckia cultivar Takii 22, wherein the Rudbeckia plants in step (b) are grown from the additional Rudbeckia plants of step (c); A method comprising: (Appendix 67) 66. A seed produced by the method described in Appendix 66, The seeds contain at least 50% alleles of the Rudbeckia plant of claim 54; A seed, wherein the Rudbeckia plant grown from said seed is cytoplasmic male sterility. (Appendix 68) A Rudbeckia plant produced by growing seeds of the Rudbeckia plant described in Appendix 67. (Appendix 69) 54. A method for introducing at least one new trait into a Rudbeckia plant of claim 54, comprising: The method comprises: (a) crossing Rudbeckia cultivar Takii 22, which is seed of the Rudbeckia plant deposited under accession number FERM BP-22428, with a Rudbeckia plant having at least one new trait to produce progeny; (b) selecting progeny containing at least one new trait; (c) crossing the progeny with Rudbeckia cultivar Takii 22 to produce backcross progeny; (d) selecting a backcross progeny that contains at least one new trait and has essentially all of the physiological and morphological characteristics of Rudbeckia cultivar Takii 22; (e) optionally, repeating steps (c) and (d) one or more times to produce a Rudbeckia plant containing at least one new trait and having essentially all of the physiological and morphological characteristics of Rudbeckia cultivar Takii 22, wherein the Rudbeckia plant in step (c) is a selected backcross progeny of step (d); A method comprising: (Appendix 70) 69. A Rudbeckia plant produced by the method described in Appendix 69. (Appendix 71) 1. A method for producing a Rudbeckia plant derived from Rudbeckia cultivar Takii 22 that includes at least one new trait, comprising: The method comprises: introducing a mutation or transgene that confers at least one trait into Rudbeckia cultivar Takii 22, which is a seed of the Rudbeckia plant deposited under accession number FERM BP-22428; A method comprising: (Appendix 72) 72. A Rudbeckia plant produced by the method described in Appendix 71. (Appendix 73) 55. A method for determining the genotype of a Rudbeckia plant according to claim 54, The method comprises: (a) obtaining a nucleic acid sample from a Rudbeckia plant described in Appendix 54; (b) detecting a polymorphism in the nucleic acid sample; A method comprising: (Appendix 74) A regenerable cell or protoplast tissue culture derived from a Rudbeckia plant described in Appendix 54. (Appendix 75) 75. The cultured tissue of claim 74, The cells or protoplasts are derived from leaves, embryos, cotyledons, hypocotyls, meristematic cells, roots, root tips, anthers, flowers, seeds or stems. Cultured tissue. (Appendix 76) 76. A Rudbeckia plant regenerated from the tissue culture described in Appendix 75. (Appendix 77) 76. A Rudbeckia plant according to claim 76, The Rudbeckia plant has cytoplasmic male sterility. Rudbeckia plant. (Appendix 78) 54. A method for vegetatively propagating a Rudbeckia plant according to claim 54, The method comprises: (a) recovering reproductive tissue from a Rudbeckia plant of Rudbeckia cultivar Takii 22, which is the seed of the Rudbeckia plant deposited under accession number FERM BP-22428; (b) culturing the tissue and obtaining proliferated shoots; (c) rooting the propagated shoots and obtaining rooted plantlets; (d) optionally growing plants from the rooted plantlets; A method comprising: (Appendix 79) 79. A Rudbeckia plantlet or plant produced by the method of claim 78, the Rudbeckia plantlet or plant has cytoplasmic male sterility; Rudbeckia plantlets or plants. <Male sterility gene> (Appendix 80) A cytoplasmic male sterility gene comprising at least one polynucleotide selected from the group consisting of (ca) to (ce) and (cf) below: (ca) any one of the following polynucleotides (ca1) to (ca7): (ca1) a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 1; (ca2) a polynucleotide consisting of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (ca1), and encoding a polypeptide that expresses cytoplasmic male sterility; (ca3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (ca1), and that causes cytoplasmic male sterility; (ca4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (ca1); (ca5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 2; (ca6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 2 in which one or several amino acids have been deleted, substituted, inserted and / or added; (ca7) a polynucleotide that expresses cytoplasmic male sterility and that consists of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 2; (cb) any one of the following polynucleotides (cb1) to (cb7): (cb1) a polynucleotide consisting of the base sequence of SEQ ID NO: 3; (cb2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cb1); (cb3) a polynucleotide that expresses cytoplasmic male sterility and that consists of a base sequence having 80% or more identity to the base sequence of (cb1); (cb4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cb1); (cb5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 4; (cb6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 4 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cb7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 4; (cc) any one of the following polynucleotides (cc1) to (cc7): (cc1) a polynucleotide consisting of the base sequence of SEQ ID NO: 5; (cc2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cc1); (cc3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (cc1), and that expresses cytoplasmic male sterility; (cc4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to the polynucleotide having the nucleotide sequence of (cc1); (cc5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 6; (cc6) A polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 6 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cc7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 6; (cd) any one of the following polynucleotides (cd1) to (cd7): (cd1) a polynucleotide consisting of the base sequence of SEQ ID NO: 7; (cd2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cd1); (cd3) a polynucleotide consisting of a base sequence having 80% or more identity to the base sequence of (cd1), which expresses cytoplasmic male sterility; (cd4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cd1); (cd5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 8; (cd6) A polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 8 in which one or several amino acids have been deleted, substituted, inserted and / or added; (cd7) a polynucleotide that expresses cytoplasmic male sterility and consists of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 8; (ce) any one of the following polynucleotides (ce1) to (ce7): (ce1) a polynucleotide consisting of the nucleotide sequence of SEQ ID NO: 9; (ce2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (ce1); (ce3) a polynucleotide that has a base sequence that is 80% or more identical to the base sequence of (ce1), and that causes cytoplasmic male sterility; (ce4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the nucleotide sequence of (ce1); (ce5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 10; (ce6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 10 in which one or several amino acids have been deleted, substituted, inserted and / or added; (ce7) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 10; (cf) any one of the following polynucleotides (cf1) to (cf7): (cf1) a polynucleotide consisting of the base sequence of SEQ ID NO: 11; (cf2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which one or several bases are deleted, substituted, inserted and / or added in the base sequence of (cf1); (cf3) a polynucleotide that expresses cytoplasmic male sterility and that consists of a base sequence having 80% or more identity to the base sequence of (cf1); (cf4) a polynucleotide that expresses cytoplasmic male sterility and that has a nucleotide sequence complementary to a polynucleotide that hybridizes under stringent conditions to the polynucleotide having the nucleotide sequence of (cf1); (cf5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 12; (cf6) a polynucleotide that expresses cytoplasmic male sterility, comprising the amino acid sequence of SEQ ID NO: 12 in which one or several amino acids have been deleted, substituted, inserted, and / or added; (cf7) A polynucleotide consisting of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 12, which polynucleotide causes expression of cytoplasmic male sterility. <Male sterility protein> (Appendix 81) A cytoplasmic male sterility protein comprising at least one polypeptide selected from the group consisting of (CA) to (CE) and (CF): (CA) any one of the following polypeptides (CA1) to (CA3): (CA1) a polypeptide consisting of the amino acid sequence of SEQ ID NO: 2; (CA2) A polypeptide that expresses cytoplasmic male sterility, consisting of an amino acid sequence in which one or several amino acids are deleted, substituted, inserted and / or added in the amino acid sequence of SEQ ID NO: 2; (CA3) A polypeptide that expresses cytoplasmic male sterility, consisting of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 2; (CB) any one of the following polypeptides (CB1) to (CB3): (CB1) a polypeptide consisting of the amino acid sequence of SEQ ID NO: 4; (CB2) A polypeptide that expresses cytoplasmic male sterility, consisting of the amino acid sequence of SEQ ID NO: 4 in which one or several amino acids have been deleted, substituted, inserted and / or added; (CB3) A polypeptide that expresses cytoplasmic male sterility, consisting of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 4; (CC) any one of the following polypeptides (CC1) to (CC3): (CC1) a polypeptide consisting of the amino acid sequence of SEQ ID NO: 6; (CC2) A polypeptide that expresses cytoplasmic male sterility, consisting of the amino acid sequence of SEQ ID NO: 6 in which one or several amino acids have been deleted, substituted, inserted and / or added; (CC3) A polypeptide that expresses cytoplasmic male sterility, consisting of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 6; (CD) any one of the following polypeptides (CD1) to (CD3): (CD1) a polypeptide consisting of the amino acid sequence of SEQ ID NO: 8; (CD2) A polypeptide that expresses cytoplasmic male sterility, consisting of the amino acid sequence of SEQ ID NO: 8 in which one or several amino acids have been deleted, substituted, inserted and / or added; (CD3) A polypeptide that expresses cytoplasmic male sterility, consisting of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 8; (CE) Any of the following polypeptides (CE1) to (CE3): (CE1) a polypeptide consisting of the amino acid sequence of SEQ ID NO: 10; (CE2) A polypeptide that expresses cytoplasmic male sterility, consisting of the amino acid sequence of SEQ ID NO: 10 in which one or several amino acids have been deleted, substituted, inserted and / or added; (CE3) A polypeptide that expresses cytoplasmic male sterility, consisting of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 10; (CF) Any of the following polypeptides (CF1) to (CF3): (CF1) a polypeptide consisting of the amino acid sequence of SEQ ID NO: 12; (CF2) A polypeptide that expresses cytoplasmic male sterility, consisting of the amino acid sequence of SEQ ID NO: 12 in which one or several amino acids have been deleted, substituted, inserted and / or added; (CF3) A polypeptide consisting of an amino acid sequence having 80% or more identity to the amino acid sequence of SEQ ID NO: 12, which expresses cytoplasmic male sterility. <Expression vector> (Appendix 82) 81. An expression vector comprising the cytoplasmic male sterility gene of claim 80. <Transformants> (Appendix 83) A transformant comprising the cytoplasmic male sterility gene of Appendix 80 or the expression vector of Appendix 82. [Industrial Applicability]

[0280] As described above, the Rudbeckia plants of the present disclosure exhibit cytoplasmic male sterility. Therefore, the present disclosure is extremely useful in, for example, the fields of breeding and agriculture.

Claims

1. A Rudbeckia plant having cytoplasmic male sterility, A Rudbeckia plant comprising the following polynucleotides (ca) to (ce) and (cf) as cytoplasmic male sterility genes: (ca) any one of the following polynucleotides (ca1) to (ca7): (ca1) a polynucleotide consisting of the base sequence of SEQ ID NO: 1; (ca2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which 1 to 67 bases are deleted, substituted, inserted and / or added in the base sequence of (ca1); (ca3) a polynucleotide consisting of a base sequence having 90% or more identity to the base sequence of (ca1), which expresses cytoplasmic male sterility; (ca4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (ca1); (ca5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 2; (ca6) a polynucleotide that expresses cytoplasmic male sterility, consisting of an amino acid sequence in which 1 to 22 amino acids are deleted, substituted, inserted and / or added in the amino acid sequence of SEQ ID NO: 2; (ca7) a polynucleotide that expresses cytoplasmic male sterility and consists of an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 2; (cb) any one of the following polynucleotides (cb1) to (cb7): (cb1) a polynucleotide consisting of the base sequence of SEQ ID NO: 3; (cb2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which 1 to 280 bases are deleted, substituted, inserted and / or added in the base sequence of (cb1); (cb3) a polynucleotide consisting of a base sequence having 90% or more identity to the base sequence of (cb1), which expresses cytoplasmic male sterility; (cb4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cb1); (cb5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 4; (cb6) a polynucleotide that expresses cytoplasmic male sterility, consisting of an amino acid sequence in which 1 to 93 amino acids are deleted, substituted, inserted and / or added in the amino acid sequence of SEQ ID NO: 4; (cb7) a polynucleotide that expresses cytoplasmic male sterility and consists of an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 4; (cc) any one of the following polynucleotides (cc1) to (cc7): (cc1) a polynucleotide consisting of the base sequence of SEQ ID NO: 5; (cc2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which 1 to 126 bases are deleted, substituted, inserted and / or added in the base sequence of (cc1); (cc3) a polynucleotide consisting of a base sequence having 90% or more identity to the base sequence of (cc1), which expresses cytoplasmic male sterility; (cc4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cc1); (cc5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 6; (cc6) a polynucleotide that expresses cytoplasmic male sterility, consisting of an amino acid sequence in which 1 to 42 amino acids are deleted, substituted, inserted and / or added in the amino acid sequence of SEQ ID NO: 6; (cc7) a polynucleotide that expresses cytoplasmic male sterility and consists of an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 6; (cd) Any of the following polynucleotides (cd1) to (cd7): (cd1) a polynucleotide consisting of the base sequence of SEQ ID NO: 7; (cd2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which 1 to 291 bases are deleted, substituted, inserted and / or added in the base sequence of (cd1); (cd3) a polynucleotide consisting of a base sequence having 90% or more identity to the base sequence of (cd1), which expresses cytoplasmic male sterility; (cd4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cd1); (cd5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 8; (cd6) A polynucleotide that expresses cytoplasmic male sterility, consisting of an amino acid sequence in which 1 to 97 amino acids are deleted, substituted, inserted and / or added in the amino acid sequence of SEQ ID NO: 8; (cd7) a polynucleotide that expresses cytoplasmic male sterility and consists of an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 8; (ce) any one of the following polynucleotides (ce1) to (ce7): (ce1) a polynucleotide consisting of the base sequence of SEQ ID NO: 9; (ce2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which 1 to 114 bases are deleted, substituted, inserted and / or added in the base sequence of (ce1); (ce3) a polynucleotide consisting of a base sequence having 90% or more identity to the base sequence of (ce1), which expresses cytoplasmic male sterility; (ce4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (ce1); (ce5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 10; (ce6) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence in which 1 to 38 amino acids are deleted, substituted, inserted and / or added in the amino acid sequence of SEQ ID NO: 10; (ce7) a polynucleotide that expresses cytoplasmic male sterility, consisting of an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 10; (cf) any one of the following polynucleotides (cf1) to (cf7): (cf1) a polynucleotide consisting of the base sequence of SEQ ID NO: 11; (cf2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which 1 to 46 bases are deleted, substituted, inserted and / or added in the base sequence of (cf1); (cf3) a polynucleotide consisting of a base sequence having 90% or more identity to the base sequence of (cf1), which expresses cytoplasmic male sterility; (cf4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cf1); (cf5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 12; (cf6) a polynucleotide that expresses cytoplasmic male sterility, consisting of an amino acid sequence in which 1 to 15 amino acids are deleted, substituted, inserted and / or added in the amino acid sequence of SEQ ID NO: 12; (cf7) A polynucleotide consisting of an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 12, which expresses cytoplasmic male sterility.

2. The Rudbeckia plant of claim 1 , which contains the cytoplasmic male sterility gene in its mitochondrial genome.

3. 3. The Rudbeckia plant according to claim 1, wherein the Rudbeckia plant is a plant grown from seeds deposited under accession number FERM BP-22428.

4. A progeny line of the Rudbeckia plant according to any one of claims 1 to 3, The progeny line exhibits cytoplasmic male sterility.

5. The progeny line of claim 4 , wherein the progeny line is a first generation hybrid line.

6. Seeds of a Rudbeckia plant according to any one of claims 1 to 3 or a progeny line according to claim 4 or 5.

7. A Rudbeckia plant according to any one of claims 1 to 3, or a part of the progeny line according to claim 4 or 5.

8. A cytoplasm contained in a Rudbeckia plant according to any one of claims 1 to 3, a progeny line according to claim 4 or 5, a seed according to claim 6, or a part according to claim 7.

9. A method for producing a cytoplasmically male sterile Rudbeckia plant, comprising the following step (a): (a) A step of crossing a Rudbeckia plant according to any one of claims 1 to 3 or a progeny line according to claim 4 or 5 with another Rudbeckia plant.

10. The method of claim 9, further comprising the following step (x) prior to step (a): (x) A step of selecting the Rudbeckia plant according to any one of claims 1 to 3 or the progeny line according to claim 4 or 5 from the test Rudbeckia plant.

11. The method according to claim 10, wherein in the step (x), a Rudbeckia plant containing the following polynucleotides (ca) to (ce) and (cf) as cytoplasmic male sterility genes is selected: (ca) any one of the following polynucleotides (ca1) to (ca7): (ca1) a polynucleotide consisting of the base sequence of SEQ ID NO: 1; (ca2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which 1 to 67 bases are deleted, substituted, inserted and / or added in the base sequence of (ca1); (ca3) a polynucleotide consisting of a base sequence having 90% or more identity to the base sequence of (ca1), which expresses cytoplasmic male sterility; (ca4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (ca1); (ca5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 2; (ca6) a polynucleotide that expresses cytoplasmic male sterility, consisting of an amino acid sequence in which 1 to 22 amino acids are deleted, substituted, inserted and / or added in the amino acid sequence of SEQ ID NO: 2; (ca7) a polynucleotide that expresses cytoplasmic male sterility and consists of an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 2; (cb) any one of the following polynucleotides (cb1) to (cb7): (cb1) a polynucleotide consisting of the base sequence of SEQ ID NO: 3; (cb2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which 1 to 280 bases are deleted, substituted, inserted and / or added in the base sequence of (cb1); (cb3) a polynucleotide consisting of a base sequence having 90% or more identity to the base sequence of (cb1), which expresses cytoplasmic male sterility; (cb4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cb1); (cb5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 4; (cb6) a polynucleotide that expresses cytoplasmic male sterility, consisting of an amino acid sequence in which 1 to 93 amino acids are deleted, substituted, inserted and / or added in the amino acid sequence of SEQ ID NO: 4; (cb7) a polynucleotide that expresses cytoplasmic male sterility and consists of an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 4; (cc) any one of the following polynucleotides (cc1) to (cc7): (cc1) a polynucleotide consisting of the base sequence of SEQ ID NO: 5; (cc2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which 1 to 126 bases are deleted, substituted, inserted and / or added in the base sequence of (cc1); (cc3) a polynucleotide consisting of a base sequence having 90% or more identity to the base sequence of (cc1), which expresses cytoplasmic male sterility; (cc4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cc1); (cc5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 6; (cc6) a polynucleotide that expresses cytoplasmic male sterility, consisting of an amino acid sequence in which 1 to 42 amino acids are deleted, substituted, inserted and / or added in the amino acid sequence of SEQ ID NO: 6; (cc7) a polynucleotide that expresses cytoplasmic male sterility and consists of an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 6; (cd) Any of the following polynucleotides (cd1) to (cd7): (cd1) a polynucleotide consisting of the base sequence of SEQ ID NO: 7; (cd2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which 1 to 291 bases are deleted, substituted, inserted and / or added in the base sequence of (cd1); (cd3) a polynucleotide consisting of a base sequence having 90% or more identity to the base sequence of (cd1), which expresses cytoplasmic male sterility; (cd4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cd1); (cd5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 8; (cd6) A polynucleotide that expresses cytoplasmic male sterility, consisting of an amino acid sequence in which 1 to 97 amino acids are deleted, substituted, inserted and / or added in the amino acid sequence of SEQ ID NO: 8; (cd7) a polynucleotide that expresses cytoplasmic male sterility and consists of an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 8; (ce) any one of the following polynucleotides (ce1) to (ce7): (ce1) a polynucleotide consisting of the base sequence of SEQ ID NO: 9; (ce2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which 1 to 114 bases are deleted, substituted, inserted and / or added in the base sequence of (ce1); (ce3) a polynucleotide consisting of a base sequence having 90% or more identity to the base sequence of (ce1), which expresses cytoplasmic male sterility; (ce4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (ce1); (ce5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 10; (ce6) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence in which 1 to 38 amino acids are deleted, substituted, inserted and / or added in the amino acid sequence of SEQ ID NO: 10; (ce7) a polynucleotide that expresses cytoplasmic male sterility, consisting of an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 10; (cf) any one of the following polynucleotides (cf1) to (cf7): (cf1) a polynucleotide consisting of the base sequence of SEQ ID NO: 11; (cf2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which 1 to 46 bases are deleted, substituted, inserted and / or added in the base sequence of (cf1); (cf3) a polynucleotide consisting of a base sequence having 90% or more identity to the base sequence of (cf1), which expresses cytoplasmic male sterility; (cf4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cf1); (cf5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 12; (cf6) a polynucleotide that expresses cytoplasmic male sterility, consisting of an amino acid sequence in which 1 to 15 amino acids are deleted, substituted, inserted and / or added in the amino acid sequence of SEQ ID NO: 12; (cf7) A polynucleotide consisting of an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 12, which expresses cytoplasmic male sterility.

12. The production method according to claim 11 , wherein in the step (x), the cytoplasmic male sterility gene is detected in the test Rudbeckia plant, and a test Rudbeckia plant containing the cytoplasmic male sterility gene is selected.

13. The production method described in claim 11 or 12, wherein in step (x), the cytoplasmic male sterility gene is detected in the mitochondrial genome of the test Rudbeckia plant, and a test Rudbeckia plant containing the cytoplasmic male sterility gene is selected.

14. A method for imparting cytoplasmic male sterility to a Rudbeckia plant, comprising the step of introducing the following polynucleotides (ca) to (ce) and (cf) as cytoplasmic male sterility genes into a target Rudbeckia plant: (ca) any one of the following polynucleotides (ca1) to (ca7): (ca1) a polynucleotide consisting of the base sequence of SEQ ID NO: 1; (ca2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which 1 to 67 bases are deleted, substituted, inserted and / or added in the base sequence of (ca1); (ca3) a polynucleotide consisting of a base sequence having 90% or more identity to the base sequence of (ca1), which expresses cytoplasmic male sterility; (ca4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (ca1); (ca5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 2; (ca6) a polynucleotide that expresses cytoplasmic male sterility, consisting of an amino acid sequence in which 1 to 22 amino acids are deleted, substituted, inserted and / or added in the amino acid sequence of SEQ ID NO: 2; (ca7) a polynucleotide that expresses cytoplasmic male sterility and consists of an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 2; (cb) any one of the following polynucleotides (cb1) to (cb7): (cb1) a polynucleotide consisting of the base sequence of SEQ ID NO: 3; (cb2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which 1 to 280 bases are deleted, substituted, inserted and / or added in the base sequence of (cb1); (cb3) a polynucleotide consisting of a base sequence having 90% or more identity to the base sequence of (cb1), which expresses cytoplasmic male sterility; (cb4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cb1); (cb5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 4; (cb6) a polynucleotide that expresses cytoplasmic male sterility, consisting of an amino acid sequence in which 1 to 93 amino acids are deleted, substituted, inserted and / or added in the amino acid sequence of SEQ ID NO: 4; (cb7) a polynucleotide that expresses cytoplasmic male sterility and consists of an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 4; (cc) any one of the following polynucleotides (cc1) to (cc7): (cc1) a polynucleotide consisting of the base sequence of SEQ ID NO: 5; (cc2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which 1 to 126 bases are deleted, substituted, inserted and / or added in the base sequence of (cc1); (cc3) a polynucleotide consisting of a base sequence having 90% or more identity to the base sequence of (cc1), which expresses cytoplasmic male sterility; (cc4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cc1); (cc5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 6; (cc6) a polynucleotide that expresses cytoplasmic male sterility, consisting of an amino acid sequence in which 1 to 42 amino acids are deleted, substituted, inserted and / or added in the amino acid sequence of SEQ ID NO: 6; (cc7) a polynucleotide that expresses cytoplasmic male sterility and consists of an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 6; (cd) Any of the following polynucleotides (cd1) to (cd7): (cd1) a polynucleotide consisting of the base sequence of SEQ ID NO: 7; (cd2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which 1 to 291 bases are deleted, substituted, inserted and / or added in the base sequence of (cd1); (cd3) a polynucleotide consisting of a base sequence having 90% or more identity to the base sequence of (cd1), which expresses cytoplasmic male sterility; (cd4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cd1); (cd5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 8; (cd6) A polynucleotide that expresses cytoplasmic male sterility, consisting of an amino acid sequence in which 1 to 97 amino acids are deleted, substituted, inserted and / or added in the amino acid sequence of SEQ ID NO: 8; (cd7) a polynucleotide that expresses cytoplasmic male sterility and consists of an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 8; (ce) any one of the following polynucleotides (ce1) to (ce7): (ce1) a polynucleotide consisting of the base sequence of SEQ ID NO: 9; (ce2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which 1 to 114 bases are deleted, substituted, inserted and / or added in the base sequence of (ce1); (ce3) a polynucleotide consisting of a base sequence having 90% or more identity to the base sequence of (ce1), which expresses cytoplasmic male sterility; (ce4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (ce1); (ce5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 10; (ce6) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence in which 1 to 38 amino acids are deleted, substituted, inserted and / or added in the amino acid sequence of SEQ ID NO: 10; (ce7) a polynucleotide that expresses cytoplasmic male sterility, consisting of an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 10; (cf) any one of the following polynucleotides (cf1) to (cf7): (cf1) a polynucleotide consisting of the base sequence of SEQ ID NO: 11; (cf2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which 1 to 46 bases are deleted, substituted, inserted and / or added in the base sequence of (cf1); (cf3) a polynucleotide consisting of a base sequence having 90% or more identity to the base sequence of (cf1), which expresses cytoplasmic male sterility; (cf4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cf1); (cf5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 12; (cf6) a polynucleotide that expresses cytoplasmic male sterility, consisting of an amino acid sequence in which 1 to 15 amino acids are deleted, substituted, inserted and / or added in the amino acid sequence of SEQ ID NO: 12; (cf7) A polynucleotide consisting of an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 12, which expresses cytoplasmic male sterility.

15. The method includes a step of conferring cytoplasmic male sterility to a target Rudbeckia plant, A method for producing a cytoplasmically male sterile Rudbeckia plant, wherein the step of imparting sterility is carried out by the imparting method according to claim 14.

16. A method for screening for cytoplasmic male sterile Rudbeckia plants, comprising a selection step of selecting, from test Rudbeckia plants, test Rudbeckia plants containing the following polynucleotides (ca) to (ce) and (cf) as cytoplasmic male sterility genes as cytoplasmic male sterile Rudbeckia plants: (ca) any one of the following polynucleotides (ca1) to (ca7): (ca1) a polynucleotide consisting of the base sequence of SEQ ID NO: 1; (ca2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which 1 to 67 bases are deleted, substituted, inserted and / or added in the base sequence of (ca1); (ca3) a polynucleotide consisting of a base sequence having 90% or more identity to the base sequence of (ca1), which expresses cytoplasmic male sterility; (ca4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (ca1); (ca5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 2; (ca6) a polynucleotide that expresses cytoplasmic male sterility, consisting of an amino acid sequence in which 1 to 22 amino acids are deleted, substituted, inserted and / or added in the amino acid sequence of SEQ ID NO: 2; (ca7) a polynucleotide that expresses cytoplasmic male sterility and consists of an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 2; (cb) any one of the following polynucleotides (cb1) to (cb7): (cb1) a polynucleotide consisting of the base sequence of SEQ ID NO: 3; (cb2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which 1 to 280 bases are deleted, substituted, inserted and / or added in the base sequence of (cb1); (cb3) a polynucleotide consisting of a base sequence having 90% or more identity to the base sequence of (cb1), which expresses cytoplasmic male sterility; (cb4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cb1); (cb5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 4; (cb6) a polynucleotide that expresses cytoplasmic male sterility, consisting of an amino acid sequence in which 1 to 93 amino acids are deleted, substituted, inserted and / or added in the amino acid sequence of SEQ ID NO: 4; (cb7) a polynucleotide that expresses cytoplasmic male sterility and consists of an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 4; (cc) any one of the following polynucleotides (cc1) to (cc7): (cc1) a polynucleotide consisting of the base sequence of SEQ ID NO: 5; (cc2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which 1 to 126 bases are deleted, substituted, inserted and / or added in the base sequence of (cc1); (cc3) a polynucleotide consisting of a base sequence having 90% or more identity to the base sequence of (cc1), which expresses cytoplasmic male sterility; (cc4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cc1); (cc5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 6; (cc6) a polynucleotide that expresses cytoplasmic male sterility, consisting of an amino acid sequence in which 1 to 42 amino acids are deleted, substituted, inserted and / or added in the amino acid sequence of SEQ ID NO: 6; (cc7) a polynucleotide that expresses cytoplasmic male sterility and consists of an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 6; (cd) Any of the following polynucleotides (cd1) to (cd7): (cd1) a polynucleotide consisting of the base sequence of SEQ ID NO: 7; (cd2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which 1 to 291 bases are deleted, substituted, inserted and / or added in the base sequence of (cd1); (cd3) a polynucleotide consisting of a base sequence having 90% or more identity to the base sequence of (cd1), which expresses cytoplasmic male sterility; (cd4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cd1); (cd5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 8; (cd6) A polynucleotide that expresses cytoplasmic male sterility, consisting of an amino acid sequence in which 1 to 97 amino acids are deleted, substituted, inserted and / or added in the amino acid sequence of SEQ ID NO: 8; (cd7) a polynucleotide that expresses cytoplasmic male sterility and consists of an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 8; (ce) any one of the following polynucleotides (ce1) to (ce7): (ce1) a polynucleotide consisting of the base sequence of SEQ ID NO: 9; (ce2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which 1 to 114 bases are deleted, substituted, inserted and / or added in the base sequence of (ce1); (ce3) a polynucleotide consisting of a base sequence having 90% or more identity to the base sequence of (ce1), which expresses cytoplasmic male sterility; (ce4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (ce1); (ce5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 10; (ce6) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence in which 1 to 38 amino acids are deleted, substituted, inserted and / or added in the amino acid sequence of SEQ ID NO: 10; (ce7) a polynucleotide that expresses cytoplasmic male sterility, consisting of an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 10; (cf) any one of the following polynucleotides (cf1) to (cf7): (cf1) a polynucleotide consisting of the base sequence of SEQ ID NO: 11; (cf2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which 1 to 46 bases are deleted, substituted, inserted and / or added in the base sequence of (cf1); (cf3) a polynucleotide consisting of a base sequence having 90% or more identity to the base sequence of (cf1), which expresses cytoplasmic male sterility; (cf4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cf1); (cf5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 12; (cf6) a polynucleotide that expresses cytoplasmic male sterility, consisting of an amino acid sequence in which 1 to 15 amino acids are deleted, substituted, inserted and / or added in the amino acid sequence of SEQ ID NO: 12; (cf7) A polynucleotide consisting of an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 12, which expresses cytoplasmic male sterility.

17. a screening step of screening test Rudbeckia plants containing a cytoplasmic male sterility gene from test Rudbeckia plants; A method for producing a cytoplasmically male sterile Rudbeckia plant, wherein the screening step is carried out by the screening method according to claim 16.

18. A method for detecting cytoplasmic male sterility in a Rudbeckia plant, comprising a detection step of detecting the following polynucleotides (ca) to (ce) and (cf) as cytoplasmic male sterility genes in a test Rudbeckia plant: (ca) any one of the following polynucleotides (ca1) to (ca7): (ca1) a polynucleotide consisting of the base sequence of SEQ ID NO: 1; (ca2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which 1 to 67 bases are deleted, substituted, inserted and / or added in the base sequence of (ca1); (ca3) a polynucleotide consisting of a base sequence having 90% or more identity to the base sequence of (ca1), which expresses cytoplasmic male sterility; (ca4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (ca1); (ca5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 2; (ca6) a polynucleotide that expresses cytoplasmic male sterility, consisting of an amino acid sequence in which 1 to 22 amino acids are deleted, substituted, inserted and / or added in the amino acid sequence of SEQ ID NO: 2; (ca7) a polynucleotide that expresses cytoplasmic male sterility and consists of an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 2; (cb) any one of the following polynucleotides (cb1) to (cb7): (cb1) a polynucleotide consisting of the base sequence of SEQ ID NO: 3; (cb2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which 1 to 280 bases are deleted, substituted, inserted and / or added in the base sequence of (cb1); (cb3) a polynucleotide consisting of a base sequence having 90% or more identity to the base sequence of (cb1), which expresses cytoplasmic male sterility; (cb4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cb1); (cb5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 4; (cb6) a polynucleotide that expresses cytoplasmic male sterility, consisting of an amino acid sequence in which 1 to 93 amino acids are deleted, substituted, inserted and / or added in the amino acid sequence of SEQ ID NO: 4; (cb7) a polynucleotide that expresses cytoplasmic male sterility and consists of an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 4; (cc) any one of the following polynucleotides (cc1) to (cc7): (cc1) a polynucleotide consisting of the base sequence of SEQ ID NO: 5; (cc2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which 1 to 126 bases are deleted, substituted, inserted and / or added in the base sequence of (cc1); (cc3) a polynucleotide consisting of a base sequence having 90% or more identity to the base sequence of (cc1), which expresses cytoplasmic male sterility; (cc4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cc1); (cc5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 6; (cc6) a polynucleotide that expresses cytoplasmic male sterility, consisting of an amino acid sequence in which 1 to 42 amino acids are deleted, substituted, inserted and / or added in the amino acid sequence of SEQ ID NO: 6; (cc7) a polynucleotide that expresses cytoplasmic male sterility and consists of an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 6; (cd) Any of the following polynucleotides (cd1) to (cd7): (cd1) a polynucleotide consisting of the base sequence of SEQ ID NO: 7; (cd2) a polynucleotide that expresses cytoplasmic male sterility, comprising a base sequence in which 1 to 291 bases are deleted, substituted, inserted and / or added in the base sequence of (cd1); (cd3) a polynucleotide consisting of a base sequence having 90% or more identity to the base sequence of (cd1), which expresses cytoplasmic male sterility; (cd4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cd1); (cd5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 8; (cd6) A polynucleotide that expresses cytoplasmic male sterility, consisting of an amino acid sequence in which 1 to 97 amino acids are deleted, substituted, inserted and / or added in the amino acid sequence of SEQ ID NO: 8; (cd7) a polynucleotide that expresses cytoplasmic male sterility and consists of an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 8; (ce) any one of the following polynucleotides (ce1) to (ce7): (ce1) a polynucleotide consisting of the base sequence of SEQ ID NO: 9; (ce2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which 1 to 114 bases are deleted, substituted, inserted and / or added in the base sequence of (ce1); (ce3) a polynucleotide consisting of a base sequence having 90% or more identity to the base sequence of (ce1), which expresses cytoplasmic male sterility; (ce4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (ce1); (ce5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 10; (ce6) a polynucleotide that expresses cytoplasmic male sterility, comprising an amino acid sequence in which 1 to 38 amino acids are deleted, substituted, inserted and / or added in the amino acid sequence of SEQ ID NO: 10; (ce7) a polynucleotide that expresses cytoplasmic male sterility, consisting of an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 10; (cf) any one of the following polynucleotides (cf1) to (cf7): (cf1) a polynucleotide consisting of the base sequence of SEQ ID NO: 11; (cf2) a polynucleotide that expresses cytoplasmic male sterility, which consists of a base sequence in which 1 to 46 bases are deleted, substituted, inserted and / or added in the base sequence of (cf1); (cf3) a polynucleotide consisting of a base sequence having 90% or more identity to the base sequence of (cf1), which expresses cytoplasmic male sterility; (cf4) a polynucleotide that expresses cytoplasmic male sterility and that has a base sequence complementary to a polynucleotide that hybridizes under stringent conditions to a polynucleotide having the base sequence of (cf1); (cf5) a polynucleotide encoding a polypeptide consisting of the amino acid sequence of SEQ ID NO: 12; (cf6) a polynucleotide that expresses cytoplasmic male sterility, consisting of an amino acid sequence in which 1 to 15 amino acids are deleted, substituted, inserted and / or added in the amino acid sequence of SEQ ID NO: 12; (cf7) A polynucleotide consisting of an amino acid sequence having 90% or more identity to the amino acid sequence of SEQ ID NO: 12, which expresses cytoplasmic male sterility.

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