WO2013080973A1 - Gène qsd1 gouvernant les dormances des graines de plantes, et utilisation de celui-ci - Google Patents

Gène qsd1 gouvernant les dormances des graines de plantes, et utilisation de celui-ci Download PDF

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WO2013080973A1
WO2013080973A1 PCT/JP2012/080631 JP2012080631W WO2013080973A1 WO 2013080973 A1 WO2013080973 A1 WO 2013080973A1 JP 2012080631 W JP2012080631 W JP 2012080631W WO 2013080973 A1 WO2013080973 A1 WO 2013080973A1
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dna
plant
seed dormancy
gene
qsd1
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Japanese (ja)
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佐藤　和広
隆夫 小松田
松本　隆
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国立大学法人岡山大学
独立行政法人農業生物資源研究所
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
    • C12Q1/6895—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for plants, fungi or algae
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
    • A01H1/04—Processes of selection involving genotypic or phenotypic markers; Methods of using phenotypic markers for selection
    • A01H1/045—Processes of selection involving genotypic or phenotypic markers; Methods of using phenotypic markers for selection using molecular markers
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
    • C12N15/8262—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield involving plant development
    • C12N15/8267—Seed dormancy, germination or sprouting
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/156—Polymorphic or mutational markers

Definitions

  • the present invention relates to the Qsd1 gene that governs the seed dormancy of plants, the determination of the degree of seed dormancy of plants using the gene, and the production of plants with altered dormancy.
  • Crod dormancy is one of the environmentally adaptive traits of wild plants, and is particularly important for wild plants in dry areas such as the Middle East, which do not germinate under poor summer conditions.
  • cultivated plants usually have low dormancy due to strong agricultural selection that shortens the period from harvesting to sowing, but if the dormancy is too shallow, the sprouting is caused by rainfall during the harvest season and the crop is fatally killed. Quality degradation.
  • the dormancy is too deep in brewing barley, it will not germinate uniformly, which will affect the production of malt.
  • dormancy of seeds is extremely important from the viewpoint of crop evolution and industry.
  • Non-patent Document 1 the homology of the genome has confirmed that a common gene with the seed dormancy gene of barley is also present in wheat. If the dormancy gene of barley is identified, more germination It is expected to be applicable to the control of dormancy in severely damaged wheat.
  • Non-Patent Document 2 Four species of seed dormancy QTLs derived from wild barley isolated in hybrid progenies by crossing between cultivated barley “Haruna Nijo” and wild barley “H602” have been identified so far (Non-Patent Document 2), one of which is Qsd1 It is. These QTLs sit on a high-density linkage map based on EST markers (Non-patent Document 3). A marker linked to Qsd1 is known (Patent Document 1).
  • the present invention has been made in view of such a situation, and an object thereof is to identify a novel gene that controls the degree of seed dormancy in plants. Another object of the present invention is to provide a method for efficiently determining the degree of seed dormancy of a plant using the identified gene. A further object of the present invention is to provide a method for efficiently producing a plant with altered seed dormancy using the identified gene.
  • Qsd1 is quantitative in character, and in particular, weakly dormant homozygous individuals and heterozygous individuals cannot be clearly distinguished by phenotype, and link the relationship between phenotype and genotype. Is difficult. Therefore, even if a high-density linkage map or linkage marker is known, it is difficult to narrow down responsible genes by a general positional cloning technique.
  • the present inventors investigated the germination rate of barley seeds that had been dormant and awakened at 25 ° C. for 5 weeks for Qsd1 , and the germination rate was 0 to 49% under the conditions, and the dormant homozygous 50% Genotypes were estimated with ⁇ 89% as heterogeneous and 90-100% as weakly dormant homozygous, and based on this estimation, attempts were made to narrow down responsible genes.
  • a large-scale population that segregates only Qsd1 BC 3 F 2 generation 910 individuals and BC 3 F 3 generation 4,792 individuals was used.
  • the barley ESTs in the Qsd1 region are extremely homologous to the rice chromosome 9 gene, the barley ESTs corresponding to the genes in the rice genome were arranged in order to confirm recombination. By carrying out such ingenuity, it was finally possible to narrow down the Qsd1 gene candidates to two candidate genes. However, it was not possible to identify which is the Qsd1 gene.
  • the present inventors identified the Qsd1 gene that controls the seed dormancy of barley, identified the single nucleotide polymorphism (SNP) correlated with the seed dormancy, and the weak seed dormancy type gene was dominant.
  • the inventors succeeded in elucidating that individuals having a heterozygous gene have a weak seed dormancy phenotype.
  • the present inventors also succeeded in identifying the corresponding wheat Qsd1 gene based on the sequence information of the barley Qsd1 gene.
  • the present inventors can efficiently determine the degree of seed dormancy of a plant and produce a plant with altered seed dormancy by using the identified Qsd1 gene. As a result, the present invention has been completed.
  • the present invention relates to the Qsd1 gene that governs the seed dormancy of plants, the determination of the degree of seed dormancy of plants using the gene, and the production of plants with modified dormancy, and more specifically, provides the following: Is.
  • DNA according to any one of the following (a) to (d), which encodes a protein having an activity of weakening a plant seed dormancy trait.
  • A DNA encoding a protein comprising the amino acid sequence set forth in SEQ ID NO: 2, 8, or 11
  • B DNA containing the coding region of the base sequence described in SEQ ID NO: 1, 3, 7, 9 or 10
  • C DNA encoding a protein comprising an amino acid sequence in which one or more amino acids are substituted, deleted, added and / or inserted in the amino acid sequence of SEQ ID NO: 2, 8 or 11
  • D DNA that hybridizes under stringent conditions with DNA comprising the nucleotide sequence set forth in SEQ ID NO: 1, 3, 7, 9 or 10
  • the DNA according to any one of (a) to (d) below, which encodes a protein having an activity that enhances a plant seed dormancy trait.
  • A DNA encoding a protein comprising the amino acid sequence set forth in SEQ ID NO: 5
  • B DNA containing the coding region of the nucleotide sequence set forth in SEQ ID NO: 4 or 6
  • C DNA encoding a protein comprising an amino acid sequence in which one or more amino acids are substituted, deleted, added, and / or inserted in the amino acid sequence of SEQ ID NO: 5
  • D DNA that hybridizes under stringent conditions with the DNA comprising the nucleotide sequence set forth in SEQ ID NO: 4 or 6 [3]
  • the DNA according to any one of the following (a) to (c), which has an activity of enhancing a plant seed dormancy trait.
  • A DNA encoding a double-stranded RNA complementary to the transcription product of DNA according to [1]
  • B DNA encoding an antisense RNA complementary to the transcription product of DNA according to [1]
  • C DNA encoding an RNA having ribozyme activity that specifically cleaves the transcript of the DNA according to [1] [4]
  • a vector comprising the DNA according to any one of [1] to [3].
  • [5] A plant cell into which the DNA according to any one of [1] to [3] is introduced.
  • a method for producing a plant in which the seed dormancy trait is weakened comprising the step of introducing the DNA according to [1] into the plant.
  • [10] A method for producing a plant with enhanced seed dormancy characteristics, characterized by suppressing the expression or function of the DNA according to [1] in a plant.
  • a method for producing a plant with enhanced seed dormancy characteristics comprising the step of introducing the DNA according to [2] or [3] into the plant.
  • An agent for weakening a plant seed dormancy trait comprising the DNA according to [1] or a vector into which the DNA is inserted.
  • a drug for enhancing the seed dormancy trait of plants comprising the DNA according to [2] or [3] or a vector into which the DNA is inserted.
  • [14] A method for determining the degree of seed dormancy in a plant, wherein the base sequence of the DNA described in [1] or [2] in the test plant is analyzed and compared with a control base sequence And how to.
  • a method for breeding a plant having a weak seed dormancy trait (A) a step of crossing a plant variety of a weak seed dormancy trait with any plant variety, (B) a step of determining the degree of seed dormancy in the individual obtained by mating in step (a) by the method according to [14], and (c) a variety determined to have weak seed dormancy Selecting the method.
  • a method for breeding a plant having a strong seed dormancy trait (A) a step of crossing a plant variety having a strong seed dormancy trait with any plant variety, (B) a step of determining the degree of seed dormancy in the individual obtained by mating in step (a) by the method according to [14], and (c) a variety determined to have strong seed dormancy Selecting the method.
  • seed dormancy means a property that does not germinate even under conditions suitable for seed germination.
  • the term “weak seed dormancy trait” means that the dormancy is maintained for a short period after the seed has matured and a high germination rate is exhibited under assay conditions suitable for germination.
  • the “strong seed dormancy trait” means that the period during which the dormancy is maintained after the seed has matured is long, and shows a low germination rate under test conditions suitable for germination. The germination rate, which is an index of dormancy strength, can be tested by the method described in Example (1).
  • a novel gene Qsd1 that controls plant seed dormancy was identified, and the position and structure of the gene on the chromosome were elucidated.
  • This provided a method for determining the degree of seed dormancy of plants targeting the Qsd1 gene, and a method for breeding plants with modified seed dormancy using the determination method.
  • a method for producing a plant with altered dormancy using the Qsd1 gene was provided.
  • FIG. 5 is an alignment diagram of barley full-length sequence NIASHv3013O02 and wheat full-length sequence RFL_Contig4246.
  • FIG. 13 is a continuation of FIG. 12.
  • FIG. 14 is a continuation of FIG. 13.
  • the present invention provides a DNA encoding a protein having an activity of weakening a plant seed dormancy trait (hereinafter referred to as “weak seed dormancy type DNA”).
  • the base sequence of Qsd1 cDNA derived from Haruna Nijo which is a weak seed dormant variety of barley, identified by the present inventors, is SEQ ID NO: 1, and the amino acid sequence of the protein encoded by the DNA is SEQ ID NO: 2. Show.
  • the base sequence of Qsd1 genomic DNA derived from Haruna Nijo is shown in SEQ ID NO: 3.
  • the base sequence of Qsd1 cDNA derived from RFL_Contig4246 in Chinese Spring which is a weak seed dormant variety of wheat, identified by the present inventors, is SEQ ID NO: 7, and the amino acid sequence encoded by the DNA is SEQ ID NO: : Shown in 8.
  • the base sequence of contig6-derived Qsd1 genomic DNA in Chinese Spring is SEQ ID NO: 9
  • the base sequence of Qsd1 cDNA extracted from contig6-derived Qsd1 genomic DNA is SEQ ID NO: 10, and the amino acid sequence encoded by these DNAs. Is shown in SEQ ID NO: 11.
  • One embodiment of the weak seed dormant DNA of the present invention is a DNA encoding a protein consisting of the amino acid sequence set forth in SEQ ID NO: 2, 8, or 11 (typically, SEQ ID NOs: 1, 3, 7, DNA containing the coding region of the base sequence described in 9 or 10.
  • a DNA encoding a protein having an activity that weakens the seed dormancy trait of a plant is encoding a protein having an activity that weakens the seed dormancy trait of a plant.
  • plurality refers to the number of amino acid modifications within the range in which the modified Qsd1 protein maintains the activity of weakening the plant seed dormancy trait, usually within 50 amino acids, preferably within 30 amino acids, Preferably, it is within 10 amino acids (eg, within 5 amino acids, within 3 amino acids, 2 amino acids).
  • weak seed dormant DNA from a specific weak seed dormant plant variety (for example, barley Haruna Nijo, wheat Chinese Spring).
  • a specific weak seed dormant plant variety for example, barley Haruna Nijo, wheat Chinese Spring
  • weak seed dormant DNA By using the base sequence information of weak seed dormant DNA, other barley varieties (eg Harbin Nijo, Russia6, cross A), other wheat varieties (eg spring love), other plant varieties (eg It is possible to obtain DNA encoding a homologous gene, which is also a weak seed dormancy type, from a wheat series such as rye.
  • the present invention relates to DNA that hybridizes under stringent conditions with Qsd1 DNA (SEQ ID NO: 1 or 3) in Haruna Nijo of Barley or Qsd1 DNA (SEQ ID NO: 7, 9, or 10) in Chinese Chinese Spring. It also includes DNA encoding a protein having an activity of weakening the plant seed dormancy trait.
  • mutant DNA or homologous DNA thus obtained encodes a protein having an activity that weakens the plant seed dormancy trait is determined by, for example, strong seed dormancy by introducing these DNAs by genetic recombination technology or crossing. It can be determined by collecting seeds from the cultivar of the mold, performing the germination test described in this example, and examining whether the germination rate is increased. If the germination rate is high, it is evaluated that it has an activity to weaken the seed dormancy trait of the plant.
  • the present invention also provides a DNA encoding a protein having an activity to enhance the seed dormancy traits of plants (hereinafter referred to as “strong seed dormancy type DNA”).
  • strong seed dormancy type DNA The base sequence of Qsd1 cDNA derived from H602, which is a barley strong seed dormant variety identified by the present inventors, is shown in SEQ ID NO: 4, and the amino acid sequence of the protein encoded by the DNA is shown in SEQ ID NO: 5.
  • the nucleotide sequence of H602-derived Qsd1 genomic DNA is shown in SEQ ID NO: 6.
  • One embodiment of the strong seed dormant DNA of the present invention is a DNA encoding a protein consisting of the amino acid sequence set forth in SEQ ID NO: 5 (typically, the base sequence code set forth in SEQ ID NO: 4 or 6). DNA containing the region).
  • the amino acid sequence of the Qsd1 protein derived from barley H602 is the amino acid sequence of the Qsd1 protein derived from Haruna Nijo, a weak seed dormant variety (SEQ ID NO: 2).
  • An amino acid substitution occurs at position 214 (see FIGS. 8 and 9). For this reason, it is considered that the function of the original Qsd1 protein is suppressed, and thereby the individual has a strong seed dormancy character.
  • the present invention comprises a protein comprising an amino acid sequence in which one or more amino acids are substituted, deleted, added and / or inserted in the amino acid sequence of the Qsd1 protein (SEQ ID NO: 5) in barley H602, It also contains DNA encoding a protein having an activity that enhances the seed dormancy traits of rice.
  • the term “plurality” refers to the number of amino acid modifications within a range in which the modified Qsd1 protein has an activity to enhance the seed dormancy trait of the plant. If the Qsd1 protein in the plant does not exhibit its original function, it is considered that it becomes a strong seed dormancy trait. Therefore, the number of amino acid modifications is essentially not limited. The modification is, for example, within 100 amino acids (50 amino acids, 30 amino acids, 10 amino acids, 5 amino acids, 3 amino acids, 2 amino acids).
  • the present invention is a DNA that hybridizes under stringent conditions with Qsd1 DNA (SEQ ID NO: 4 or 6) in barley H602, and encodes a protein having an activity that enhances the plant seed dormancy trait Contains DNA.
  • mutant DNA or homologous DNA obtained in this way encodes a protein having an activity that enhances the seed dormancy trait of the plant is, for example, recombining the Qsd1 gene of the weak seed dormancy variety with the DNA, It can be determined by creating a plant that holds the DNA homologously, collecting seeds from the plant, performing the germination test described in this example, and testing whether the germination rate is low. it can. If the germination rate is low, it is evaluated that the plant has an activity to enhance the seed dormancy trait of the plant.
  • the weak seed dormancy type DNA of the present invention is an agent for weakening a plant seed dormancy trait in the sense that introduction thereof can weaken the plant seed dormancy trait
  • the strong seed dormancy type DNA of the present invention is a drug for enhancing the seed dormancy of a plant in the sense that its introduction can enhance the plant seed dormancy trait.
  • artificial mutations introduced into DNA to produce the above-mentioned mutant DNA are, for example, site-directed mutagenesis (Kramer, W. & Fritz, HJ., Methods Enzymol). , 154: 350-367, 1987).
  • hybridization technology Southern, E. M., Journal of Molecular Biology, 98: 503, 1975
  • PCR polymerase chain reaction
  • Saiki polymerase chain reaction
  • stringent hybridization conditions include 6M urea, 0.4% SDS, 0.5xSSC conditions, or equivalent stringency hybridization conditions.
  • Isolation of DNA with higher homology can be expected by using conditions with higher stringency, for example, 6M urea, 0.4% SDS, and 0.1xSSC.
  • the isolated DNA is at least 50% or more, more preferably 70% or more, more preferably 90% or more (for example, 95%, 96%, 97%, 98%, 99%) at the nucleic acid level or amino acid sequence level. And the like).
  • Sequence homology can be determined using BLASTN (nucleic acid level) and BLASTX (amino acid level) programs (Altschul et al. J. Mol. Biol., 215: 403-410, 1990). The program is based on the algorithm BLAST (Proc. Natl. Acad. Sci.
  • the DNA encoding the Qsd1 protein of the present invention is not particularly limited in its form, and includes cDNA, genomic DNA, and chemically synthesized DNA. Preparation of genomic DNA and cDNA can be performed by those skilled in the art using conventional means.
  • genomic DNA for example, genomic DNA is extracted from a plant, a genomic library (a vector such as a plasmid, phage, cosmid, BAC, PAC, etc. can be used), expanded, and Qsd1 gene (for example, Prepared by performing colony hybridization or plaque hybridization using a probe prepared on the basis of the nucleotide sequence of the DNA of SEQ ID NO: 1,3,4,6,7,9,10) It is possible.
  • cDNA is synthesized based on mRNA extracted from a plant, inserted into a vector such as ⁇ ZAP to create a cDNA library, developed, and colony hybridization in the same manner as described above.
  • a vector such as ⁇ ZAP
  • it can be prepared by performing plaque hybridization or by performing PCR.
  • the present invention also provides DNA used to suppress the expression of weak seed dormancy type Qsd1 gene in plants.
  • the DNA used to suppress the expression of the weak seed dormancy type Qsd1 gene in the plant is a drug for enhancing the plant seed dormancy trait.
  • “suppression of Qsd1 gene expression” includes both suppression of gene transcription and suppression of protein translation. Further, “suppression of expression” includes not only complete cessation of expression but also decrease of expression.
  • a wheat series (Triticeae) plant such as barley, wheat, and rye is preferred, and barley and wheat are particularly preferred.
  • RNAi RNA interference
  • Dicer RNaseIII-like nuclease
  • siRNA short interference RNA
  • a specific protein binds to this siRNA to form a nuclease complex (RISC: RNA-induced silencing complex).
  • RISC RNA-induced silencing complex
  • This complex recognizes and binds to the same sequence as siRNA, and cleaves the transcript (mRNA) of the target gene at the center of the siRNA by RNaseIII-like enzyme activity.
  • the antisense strand of siRNA binds to mRNA and acts as a primer for RNA-dependent RNA polymerase (RsRP) to synthesize dsRNA.
  • RsRP RNA-dependent RNA polymerase
  • the DNA encoding the dsRNA of the present invention includes an antisense DNA encoding an antisense RNA for any region of a target gene transcription product (mRNA), and a sense DNA encoding a sense RNA for any region of the mRNA
  • antisense RNA and sense RNA can be expressed from the antisense DNA and the sense DNA, respectively.
  • dsRNA can be produced from these antisense RNA and sense RNA.
  • the dsRNA expression system of the present invention is held in a vector or the like, there are a case where antisense RNA and sense RNA are expressed from the same vector, and a case where antisense RNA and sense RNA are expressed from different vectors, respectively. is there.
  • the antisense RNA and sense RNA are expressed from the same vector.
  • an antisense RNA expression cassette in which a promoter capable of expressing a short RNA such as polIII is linked upstream of the antisense DNA and the sense DNA.
  • sense RNA expression cassettes are constructed, and these cassettes are inserted into the vector in the same direction or in the opposite direction.
  • an expression system in which antisense DNA and sense DNA are arranged in opposite directions so as to face each other on different strands.
  • one double-stranded DNA in which an antisense RNA coding strand and a sense RNA coding strand are paired is provided, and antisense RNA and sense RNA are separated from each strand on both sides.
  • a promoter is provided oppositely so that it can be expressed.
  • a terminator is added to the 3 'end of each strand (antisense RNA coding strand, sense RNA coding strand). It is preferable to provide.
  • this terminator a sequence in which four or more A (adenine) bases are continued can be used.
  • the two promoter types are preferably different.
  • antisense RNA expression in which a promoter capable of expressing a short RNA such as polIII is linked upstream of antisense DNA and sense DNA, respectively.
  • a cassette and a sense RNA expression cassette are constructed, and these cassettes are held in different vectors.
  • the dsRNA used in the present invention is preferably siRNA.
  • siRNA means a double-stranded RNA consisting of short strands in a range that is not toxic in cells.
  • the chain length is not particularly limited as long as the expression of the target Qsd1 gene can be suppressed and it does not show toxicity.
  • the dsRNA chain length is, for example, 15 to 49 base pairs, preferably 15 to 35 base pairs, and more preferably 21 to 30 base pairs.
  • an appropriate sequence preferably an intron sequence
  • a double-stranded RNA having a hairpin structure preferably an intron sequence
  • hpRNA self-complementary 'hairpin' RNA
  • the DNA encoding the dsRNA of the present invention need not be completely identical to the base sequence of the target Qsd1 gene, but is at least 70% or more, preferably 80% or more, more preferably 90% or more (for example, 95%, 96%, 97%, 98%, 99% or more). Sequence identity can be determined by the technique described above (BLAST program).
  • the part of the double-stranded RNA in which the RNAs in dsRNA are paired is not limited to a perfect pair, but mismatch (corresponding base is not complementary), bulge (no base corresponding to one strand) ) Or the like may include an unpaired portion.
  • both bulges and mismatches may be included in the double-stranded RNA region where RNAs in dsRNA pair with each other.
  • Another embodiment of the DNA used for suppressing the expression of the weak seed dormancy type Qsd1 gene in the plant is a DNA encoding an antisense RNA complementary to the above-mentioned transcript of the weak seed dormancy type DNA of the present invention (Antisense DNA).
  • Antisense DNA suppresses target gene expression by inhibiting transcription initiation by triplex formation, suppressing transcription by hybridization with a site where an open loop structure is locally created by RNA polymerase, Inhibition of transcription by hybridization with certain RNA, suppression of splicing by hybridization at the junction of intron and exon, suppression of splicing by hybridization with spliceosome formation site, suppression of transition from nucleus to cytoplasm by hybridization with mRNA , Splicing suppression by hybridization with capping site and poly (A) addition site, translation initiation suppression by hybridization with translation initiation factor binding site, translation suppression by hybridization with ribosome binding site near initiation codon, translation of mRNA Area or poly Outgrowth inhibitory peptide chains by the formation of a hybrid with over arm binding sites, and the like gene silencing and the like by hybridization with interaction site between a nucleic acid and protein.
  • the antisense DNA used in the present invention may suppress the expression of the target Qsd1 gene by any of the actions described above.
  • an antisense sequence complementary to the untranslated region near the 5 ′ end of the mRNA of the target gene is designed, it will be effective for inhibiting translation of the gene.
  • sequences complementary to the coding region or the 3 ′ untranslated region can also be used.
  • a DNA containing an antisense sequence of a non-translated region as well as a translated region of a gene is also included in the antisense DNA used in the present invention.
  • the antisense DNA to be used is linked downstream of a suitable promoter, and preferably a sequence containing a transcription termination signal is linked on the 3 ′ side.
  • Antisense DNA is a phosphorothioate method (Stein, Nucleic Acids) based on the sequence information of the weak seed dormant DNA of the present invention (for example, DNA comprising the base sequence described in SEQ ID NO: 1, 7 or 10). Res., 16: 3209-3221, 1988).
  • the prepared DNA can be introduced into plants by a known method described later.
  • the sequence of the antisense DNA is preferably complementary to the endogenous weak seed dormant Qsd1 gene transcript of the plant, but is not completely complementary as long as it can effectively inhibit gene expression. May be.
  • the transcribed RNA preferably has a complementarity of 90% or more (eg, 95%, 96%, 97%, 98%, 99% or more) to the transcript of the target gene.
  • the length of the antisense DNA is at least 15 bases or more, preferably 100 bases or more, more preferably 500 bases or more.
  • the length of the antisense DNA used is shorter than 5 kb, preferably shorter than 2.5 kb.
  • RNA having a ribozyme activity that specifically cleaves the transcript of the weak seed dormant DNA of the present invention.
  • DNA Some ribozymes have a group I intron type or a size of 400 nucleotides or more like M1RNA contained in RNaseP, but some have an active domain of about 40 nucleotides called hammerhead type or hairpin type ( Makoto Koizumi and Eiko Otsuka, Protein Nucleic Acid Enzymes, 35: 2191, 1990).
  • the self-cleaving domain of hammerhead ribozyme cleaves 3 ′ of C15 of G13U14C15, but it is important for U14 to form a base pair with A at position 9, and the base at position 15 is In addition to C, it is shown that it can also be cut by A or U (Koizumi et. Al., FEBS Lett. 228: 225, 1988).
  • the ribozyme substrate binding site is designed to be complementary to the RNA sequence near the target site, it is possible to create a restriction enzyme-like RNA-cleaving ribozyme that recognizes the UC, UU, or UA sequence in the target RNA.
  • Hairpin ribozymes are also useful for the purposes of the present invention. Hairpin ribozymes are found, for example, in the minus strand of satellite RNA of tobacco ring spot virus (Buzayan, Nature 323: 349, 1986). It has been shown that this ribozyme can also be designed to cause target-specific RNA cleavage (Kikuchi and Sasaki, Nucleic Acids Res. 19: 6751, 1992, Hiroshi Kikuchi, Chemistry and Biology 30: 112, 1992). A ribozyme designed to cleave the target is linked to a promoter such as the cauliflower mosaic virus 35S promoter and a transcription termination sequence so that it is transcribed in plant cells.
  • a promoter such as the cauliflower mosaic virus 35S promoter and a transcription termination sequence
  • Such structural units can be arranged in tandem so that multiple sites in the target gene can be cleaved to further increase the effect (Yuyama et al., Biochem. Biophys. Res. Commun. 186: 1271, 1992 ).
  • the transcription product of the target Qsd1 gene can be specifically cleaved to suppress the expression of the gene.
  • the present invention also includes a vector containing the DNA of the present invention (weak seed dormancy type DNA, strong seed dormancy type DNA, DNA for suppressing the expression of the Qsd1 gene), the DNA of the present invention or the same.
  • a plant cell into which a vector is introduced a plant body containing the cell, a plant body which is a descendant or clone of the plant body, and a propagation material of these plant bodies.
  • the vector of the present invention is not particularly limited as long as the inserted gene can be expressed in plant cells.
  • the vector of the present invention may contain a promoter for constitutively or inducibly expressing the DNA of the present invention.
  • promoters for constant expression include cauliflower mosaic virus 35S promoter, rice actin promoter, maize ubiquitin promoter, and the like.
  • promoters for inducible expression are known to be expressed by external factors such as infection and invasion of filamentous fungi, bacteria and viruses, low temperature, high temperature, drying, UV irradiation, and spraying of specific compounds. The promoter etc. which are mentioned are mentioned.
  • promoters examples include rice chitinase gene promoters expressed by infection and invasion of filamentous fungi, bacteria and viruses, promoters of tobacco PR protein genes, rice lip19 gene promoters induced by low temperatures, Induced rice hsp80 and hsp72 gene promoters, Arabidopsis thaliana rab16 gene promoter induced by drying, Parsley chalcone synthase gene promoter induced by UV irradiation, corn induced under anaerobic conditions Examples include the promoter of alcohol dehydrogenase gene.
  • the rice chitinase gene promoter and tobacco PR protein gene promoter are also induced by specific compounds such as salicylic acid, and rab16 is also induced by spraying the plant hormone abscisic acid.
  • the plant derived from the plant cell into which the vector of the present invention is introduced is not particularly limited, but wheat series (Triticeae) plants such as barley, wheat and rye are preferred, and barley and wheat are particularly preferred.
  • the plant cells of the present invention include cultured cells as well as cells in the plant body. Also included are various forms of plant cells, such as suspension culture cells, protoplasts, leaf sections, callus, immature embryos, pollen and the like.
  • various methods known to those skilled in the art such as polyethylene glycol method, electroporation (electroporation), Agrobacterium-mediated method, and particle gun method can be used.
  • Regeneration of plant bodies from transformed plant cells can be performed by methods known to those skilled in the art depending on the type of plant cells.
  • methods for creating transgenic plants for barley include Tingay et al. (Tingay S. et al. Plant J. 11: 1369-1376, 1997), Murray et al. (Murray F et al. Plant Cell Report 22: 397). -402, 2004), and Travelalla et al. (Travalla S et al. Plant Cell Report 23: 780-789, 2005).
  • a technique for producing a transformed plant body in wheat for example, a method for introducing a gene into a wheat seed immature embryo using a particle gun to regenerate the plant body (Japanese Patent Laid-Open No. 2008-212048) was described. A method can be mentioned. For other plants, a known method for regenerating a plant can be used.
  • the present invention includes a plant cell into which the DNA of the present invention is introduced, a plant containing the cell, a progeny and clone of the plant, and a propagation material for the plant, its progeny and clone.
  • the present invention also provides a method for producing a plant having a modified seed dormancy trait.
  • the plant for modifying the seed dormancy trait according to the method of the present invention is not particularly limited, but wheat series (Triticeae) plants such as barley, wheat and rye are preferred, and barley and wheat are particularly preferred.
  • One embodiment of the method of the present invention is a method for producing a plant with a weakened seed dormancy character, comprising the step of introducing the weak seed dormancy type DNA of the present invention into a plant.
  • “weakening” the seed dormancy traits of plants not only weakens the seed dormancy traits of varieties having strong seed dormancy traits, but already has certain weak seed dormancy traits. It also includes further reducing the seed dormancy of varieties having
  • the plant has a seed dormancy trait comprising a step of suppressing the expression or function of the weak seed dormancy type DNA (weak seed dormancy type Qsd1 gene) of the present invention. It is an enhanced plant production method.
  • “increasing” a seed dormancy trait of a plant not only enhances the seed dormancy trait of a variety having a weak seed dormancy trait, but also already has a certain strong seed dormancy trait. It is intended to include further increasing the seed dormancy of varieties having
  • Inhibition of the expression or function of the weak seed dormancy type DNA of the present invention in plants can be carried out by introducing the above-described strong seed dormancy type DNA of the present invention into plants. Since the strong seed dormancy trait in plants is dominated by a single recessive gene, in order to impart a strong seed dormancy trait to a plant, both of the Qsd1 alleles in an individual are usually strongly seed dormant DNA. It is necessary to. Thereby, only strong seed dormancy type DNA is expressed in an individual, and the plant seed dormancy trait can be strengthened.
  • Introduction of the strong seed dormant DNA of the present invention into a plant chromosome can be performed by, for example, mating or homologous recombination.
  • a specific DNA sequence may be introduced into weak seed dormancy type DNA on the plant chromosome to destroy its function.
  • suppression of the expression or function of the weak seed dormancy type DNA of the present invention in a plant can be achieved by using DNA for suppressing the expression of the Qsd1 gene of the present invention in the plant (for example, DNA encoding dsRNA, antisense DNA, It can be carried out by introducing a DNA encoding an RNA having ribozyme activity.
  • DNA for suppressing the expression of the Qsd1 gene of the present invention in the plant for example, DNA encoding dsRNA, antisense DNA, It can be carried out by introducing a DNA encoding an RNA having ribozyme activity.
  • the weak seed dormant DNA of the present invention In order to suppress the expression or function of the weak seed dormant DNA of the present invention in plants, for example, it binds to a drug that suppresses the expression of weak seed dormant DNA or a weak seed dormant DNA translation product, and its function is reduced.
  • a suppressive drug is also conceivable.
  • the present invention also provides a method for determining the degree of seed dormancy of a plant.
  • the plant for determining the degree of seed dormancy by the method of the present invention is not particularly limited, but wheat series (Triticeae) plants such as barley, wheat and rye are preferred, and barley and wheat are particularly preferred.
  • One embodiment of the determination method of the present invention is a method characterized by analyzing the base sequence of the Qsd1 gene in a plant and comparing it with a base sequence of a control.
  • an amplification product obtained by amplifying the Qsd1 gene by PCR can be used.
  • primers used are not limited as long as it can specifically amplify the Qsd1 gene, Qsd1 gene sequence information (e.g., SEQ ID NO: 1,3,4,6,7, 9,10) can be designed as appropriate.
  • a specific base sequence of the Qsd1 gene can be amplified by appropriately combining designed primers.
  • Control nucleotide sequence as compared to the nucleotide sequence of Qsd1 gene in the test plants are typically nucleotide sequence of Qsd1 gene in the weak dormancy type variety or strong dormancy type cultivars.
  • the amino acid sequence of the Qsd1 protein derived from H602, which is a strong seed dormant variety of barley (SEQ ID NO: 5), is Qsd1 derived from Haruna Nijo, a weak seed dormant variety.
  • an amino acid substitution F ⁇ L occurs at position 214 (see FIGS. 8 and 9).
  • the 214th amino acid is a preferable index for such evaluation.
  • Whether the base sequence of the Qsd1 gene in the test plant is different from the base sequence of the control can be indirectly analyzed by various methods other than the determination of the direct base sequence described above.
  • methods include RFLP method using restriction fragment length polymorphism / RFLP, PCR-RFLP method, PCR-SSCP (single-strand conformation polymorphism, single chain higher order structure) Polymorphism) method, denaturant gradient gel electrophoresis (DGGE) method, allele specific oligonucleotide (ASO) hybridization method, and ribonuclease A mismatch cleavage method.
  • DNA can be prepared from a test plant using a conventional method, for example, CTAB method.
  • a conventional method for example, CTAB method.
  • plant seeds, seedlings, and grown plants can be used.
  • the base sequence can be determined by a conventional method such as the dideoxy method or the Maxam-Gilbert method. In determining the base sequence, a commercially available sequence kit and sequencer can be used.
  • the present invention also provides a method for breeding a plant having a modified seed dormancy trait.
  • the plant to be bred by the method of the present invention is not particularly limited, but wheat series (Triticeae) plants such as barley, wheat and rye are preferred, and barley and wheat are particularly preferred.
  • One aspect of the breeding method of the present invention is a method of breeding a plant having a strong seed dormancy trait, and (a) a step of crossing a plant variety having a strong seed dormancy trait with any plant variety, ( b) a step of determining the degree of seed dormancy in the individual obtained by the mating by the determination method of the present invention, and (c) selecting a variety determined to have strong seed dormancy.
  • Examples of “arbitrary plant varieties” to be crossed with plant varieties with strong seed dormancy traits include, for example, varieties with weak seed dormancy traits, varieties with weak seed dormancy traits, and strong seed dormancy traits Examples include, but are not limited to, individuals obtained by crossing with cultivars.
  • Another aspect of the breeding method of the present invention is a method of breeding a plant having a weak seed dormancy trait, and (a) a step of crossing a plant variety having a weak seed dormancy trait with any plant variety , (B) determining the degree of seed dormancy in the individual obtained by mating by the determination method of the present invention, and (c) selecting a variety determined to have weak seed dormancy. Including.
  • Examples of “arbitrary plant varieties” to be bred with plant varieties with weak seed dormancy traits include, for example, varieties with strong seed dormancy traits, varieties with strong seed dormancy traits, and weak seed dormancy traits Examples include, but are not limited to, individuals obtained by crossing with cultivars.
  • the stored seeds were awakened by dormancy at 25 ° C., then sown in a petri dish to absorb water, and sprouting was induced at 25 ° C. Then, the dormancy was evaluated by the germination rate (rooting rate) on the fourth day after sowing.
  • -Wheat- Seeds that have reached physiological ripening on the 60th day after flowering are harvested, then sown in a petri dish, induced to germinate at 20 ° C, and the dormancy is evaluated by the germination rate on the 7th day after sowing. be able to.
  • the genotype can be estimated with a germination rate of 0-39% as strongly dormant homo, 40-59% hetero, and 60-100% weak dormant homo (Nakamura S. et al., Plant Cell. 2011 Sep; 23 (9): 3215-29. See Figure 5A and its experimental method).
  • gene 1 When the gene sequences of Haruna Nijo and wild barley were translated into amino acids, gene 1 showed two SNPs in exons but no amino acid substitutions. On the other hand, gene 2 had four SNPs with amino acid substitutions, and was present on the putative domain (FIGS. 6 and 9). This fact suggests that the true identity of the Qsd1 gene involved in seed dormancy is gene 2, and that SNP with nonsynonymous substitution is a mutation that governs this trait.
  • Non-patent Document 2 we analyzed dormancy (d) and non-dormancy (nd) alleles of 9 populations (Non-patent Document 2) where Qsd1 was identified by analyzing seed dormancy of barley (Fig. 7). Comparison with the base of the SNP of gene 2. As a result, the allele of the non-synonymous SNP located at the most 5 ′ end of gene 2 and the allele of Qsd1 were completely matched, so it was determined that this SNP controls dormancy (FIG. 8).
  • RFL_Contig4246 GenBank: AK333743.1
  • evalue 0
  • SEQ ID NO: 12 This sequence is derived from the wheat variety Chinese Spring.
  • Primer creation Using the software Primer 3, three pairs of primers (Contig4246-1L and Contig4246-1R, Contig4246-2L and Contig4246-2R, Contig4246-2R, Contig4246-3L and the primer for selecting a BAC clone from the sequence of RFL_Contig4246 Contig4246-3R) was created.
  • the primer sequences are as follows.
  • Contig4246-1_L GTGACTCTTTGCCCCAACAT (SEQ ID NO: 13)
  • Contig4246-1_R CCTGTGGCTTGTGTAGCTGA (SEQ ID NO: 14)
  • Contig4246-2_L GAGATTCGCAAAGTGGCTTC (SEQ ID NO: 15)
  • Contig4246-2_R GAAGATGCACATCAGCTTCG (SEQ ID NO: 16)
  • Contig4246-3_R AATGGACGCCGAGTATAACG (SEQ ID NO: 18)
  • the base sequence of Qsd1 cDNA derived from RFL_Contig4246 in Chinese Spring obtained by the above analysis is shown in SEQ ID NO: 7, and the amino acid sequence encoded by the DNA is shown in SEQ ID NO: 8.
  • the base sequence of contig6-derived Qsd1 genomic DNA in Chinese Spring is SEQ ID NO: 9
  • the base sequence of Qsd1 cDNA extracted from contig6-derived Qsd1 genomic DNA is SEQ ID NO: 10
  • the amino acid sequence encoded by these DNAs Is shown in SEQ ID NO: 11.
  • a novel gene Qsd1 that governs the degree of seed dormancy of plants is identified, and using the identified Qsd1 gene, determination of the degree of seed dormancy of plants and seed dormancy can be achieved. Production and breeding of modified plants became possible.
  • the determination of the degree of seed dormancy in the present invention is targeted to the gene that controls it, and can be carried out using an individual (for example, seed) in the early stage of growth. For this reason, if the determination method of the present invention is used, it is possible to breed varieties with modified seed dormancy specifically and efficiently.
  • Weakly dormant plants produced and bred in this way can ensure sufficient and early germination individuals when dormancy is deep and germination is significantly delayed, or when germination is delayed due to low precipitation in dry areas Useful in terms. Strong seed dormancy plants are useful in that they can prevent seed quality degradation due to ear germination in areas with heavy rainfall. In addition, by controlling the seed dormancy, it is possible to promote uniform germination in the brewing barley, thereby enabling efficient malt production. Therefore, the present invention can greatly contribute to the improvement of the yield of crops and the reduction of economic loss due to the deterioration of quality.

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Abstract

La présente invention concerne un gène Qsd1 qui gouverne les dormances des graines d'orge et qui peut être identifié successivement par une technique de clonage positionnel. Un polymorphisme de simple nucléotide (SNP) qui est corrélé aux dormances des graines mentionnées ci-dessus et est situé dans le gène peut également être spécifié successivement, et il peut être observé successivement qu'un individu dans lequel un gène faiblement inducteur de dormance de graine est dominant et le gène est hétérogène a un génotype faiblement inducteur de dormance de graine. De plus, sur la base des informations de séquence sur le gène Qsd1 d'orge, le gène Qsd1 correspondant de blé peut également être identifié successivement. Ensuite, il est observé que la détermination des degrés de dormance des graines d'une plante et la production d'une plante ayant des dormances des graines modifiées peut être effectuée avec une efficacité élevée en utilisant le gène identifié.
PCT/JP2012/080631 2011-11-28 2012-11-27 Gène qsd1 gouvernant les dormances des graines de plantes, et utilisation de celui-ci WO2013080973A1 (fr)

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JP4298538B2 (ja) * 2004-02-17 2009-07-22 独立行政法人科学技術振興機構 休眠性に関与する遺伝子座に連鎖する遺伝マーカーおよびその利用

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP4298538B2 (ja) * 2004-02-17 2009-07-22 独立行政法人科学技術振興機構 休眠性に関与する遺伝子座に連鎖する遺伝マーカーおよびその利用

Non-Patent Citations (11)

* Cited by examiner, † Cited by third party
Title
DATABASE GENBANK 20 May 2011 (2011-05-20), MATSUMOTO,T. ET AL.: "Hordeum vulgare subsp. vulgare mRNA for predicted protein, complete cds, clone: NIASHv3013002", XP003031952, accession no. K372829.1 *
K. SATO ET AL.: "A high-density transcript linkage map of barley derived from a single population", HEREDITY, vol. 103, 2009, pages 110 - 117, XP055073821 *
KAZUHIDE RIKIISHI ET AL.: "Characterization of a novel wheat (Triticum aestivum L.) mutant with reduced seed dormancy", JOURNAL OF CEREAL SCIENCE, vol. 51, no. 3, 2010, pages 292 - 298, XP027058785 *
KAZUHIRO SATO ET AL.: "Genetic analysis of seed dormancy QTL in barley", BREEDING SCIENCE, vol. 59, no. 5, 2009, pages 645 - 650, XP055073816 *
KAZUHIRO SATO ET AL.: "Omugi Shushi Kyuminsei Qsd1 no Dotei", BREEDING RESEARCH, vol. 14, 29 March 2012 (2012-03-29), pages 92 *
KAZUHIRO SATO ET AL.: "Omugi Shushi Kyuminsei QTL ''Qsd1'' no Seimitsu Mapping", BREEDING RESEARCH, vol. 12, 24 September 2010 (2010-09-24), pages 128 *
KIYOSUMI HORI ET AL.: "Barley EST Markers Enhance Map Saturation and QTL Mapping in Diploid Wheat", BREEDING SCIENCE, vol. 57, no. 1, 2007, pages 39 - 45, XP055073819 *
KIYOSUMI HORI ET AL.: "Detection of seed dormancy QTL in multiple mapping populations derived from crosses involving novel barley germplasm", THEORETICAL AND APPLIED GENETICS, vol. 115, no. 6, 2007, pages 869 - 876, XP019535857 *
SHINGO NAKAMURA ET AL.: "Mapping a diploid wheat abscisic acid 8'-hydroxylase homologue in the seed dormancy QTL region on chromosome 5Am", EUPHYTICA, vol. 171, no. 1, 2010, pages 111 - 120, XP019768732 *
TAKASHI MATSUMOTO ET AL.: "Comprehensive Sequence Analysis of 24,783 Barley Full-Length cDNAs Derived from 12 Clone Libraries", PLANT PHYSIOLOGY, vol. 156, no. 1, May 2011 (2011-05-01), pages 20 - 28, XP002670706 *
XING-YOU GU ET AL.: "The qSD12 underlying gene promotes abscisic acid accumulation in early developing seeds to induce primary dormancy in rice", PLANT MOLECULAR BIOLOGY, vol. 73, no. 1-2, 2010, pages 97 - 104, XP019794081 *

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