Nothing Special   »   [go: up one dir, main page]

WO2020156406A1 - Gène haploïde induit de blé et application associée - Google Patents

Gène haploïde induit de blé et application associée Download PDF

Info

Publication number
WO2020156406A1
WO2020156406A1 PCT/CN2020/073688 CN2020073688W WO2020156406A1 WO 2020156406 A1 WO2020156406 A1 WO 2020156406A1 CN 2020073688 W CN2020073688 W CN 2020073688W WO 2020156406 A1 WO2020156406 A1 WO 2020156406A1
Authority
WO
WIPO (PCT)
Prior art keywords
wheat
sequence
pla
gene
haploid
Prior art date
Application number
PCT/CN2020/073688
Other languages
English (en)
Chinese (zh)
Inventor
陈绍江
刘晨旭
钟裕
祁晓龙
刘宗凯
Original Assignee
中国农业大学
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 中国农业大学 filed Critical 中国农业大学
Publication of WO2020156406A1 publication Critical patent/WO2020156406A1/fr

Links

Images

Classifications

    • 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
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/16—Hydrolases (3) acting on ester bonds (3.1)
    • 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
    • A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
    • 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)
    • C12N15/8216—Methods for controlling, regulating or enhancing expression of transgenes in plant cells
    • C12N15/8218—Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS]
    • 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
    • C12N15/8287—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for fertility modification, e.g. apomixis

Definitions

  • the invention relates to the field of biotechnology, in particular to wheat haploid induction genes and applications thereof.
  • wheat is the main food crop in the world. Maintaining a steady increase in wheat yield and continuous improvement of quality is still an important task for wheat breeding.
  • wheat is a typical self-pollinated crop, and the varieties used in production are also genetically homozygous pure line materials. Therefore, although the breeding of new wheat varieties does not require pollination every generation, it still needs to be self-purified generation by generation. After 8 generations of natural selfing or above, excellent pure line materials can be obtained. Although artificial pollination is not required, since most wheat materials can only be planted one generation per year, it still consumes a lot of time and energy.
  • the genome of wheat is heterohexaploid, and the relatively complex characteristics of the genome significantly increase the difficulty of wheat breeding.
  • haploid sports seeding method can significantly improve breeding efficiency, and has been widely used in corn and other crops. Therefore, it is of great significance to establish a technical system of wheat haploid seed.
  • Predecessors explored a variety of methods to produce wheat haploids, such as gametophyte culture, distant hybridization, and apomixis.
  • gametophyte culture and distant hybridization methods still require a lot of tedious operations, or use corn pollen, which is not efficient and is greatly affected by the material background, making it difficult to use on a large scale.
  • the haploid induction method based on induced genes has been successfully applied in maize on a large scale, and its efficiency can reach more than 10%. It has become the main method for the selection of maize backbone inbred lines. Therefore, if the method of haploid induction in maize can be applied to wheat haploid seed, the efficiency of wheat haploid seed can be greatly improved.
  • the technical problem to be solved by the present invention is how to improve the efficiency of wheat haplotype.
  • the present invention first provides a method for preparing a wheat female parent haploid induction line.
  • the preparation method of the wheat maternal haploid induction line provided by the present invention includes the following steps: silencing or inhibiting the expression and/or activity of the PLA gene in the target wheat genome or knocking out the PLA gene to obtain transgenic wheat, which is the wheat maternal single Ploidy induction line;
  • the PLA gene is the PLA-A gene in the wheat A genome and/or the PLA-B gene in the wheat B genome and/or the PLA-D gene in the wheat D genome.
  • the genomic sequences of the PLA-A gene, the PLA-B gene and the PLA-D gene are as shown in sequence 1, sequence 4 and sequence 7 in the sequence listing, respectively.
  • the silencing or suppressing the expression and/or activity of the PLA gene in the target wheat genome or knocking out the PLA gene is to mutate the PLA gene in the target wheat genome to reduce the PLA gene expression in the target wheat genome or to reduce the PLA gene in the target wheat genome.
  • Gene deletion mutation or insertion mutation or base substitution is to be used.
  • the method for causing deletion mutation, insertion mutation or base substitution in the PLA gene in the target wheat genome is CRISPR/Cas9.
  • the target sequence of the CRISPR/Cas9 is 905-923 of sequence 1 and 939-957 of sequence 7.
  • the target sequence of the CRISPR/Cas9 is the 699-718th position of sequence 1.
  • the silencing or suppressing the expression and/or activity of the PLA gene in the target wheat genome or knocking out the PLA gene is performed by introducing the substance that knocks out the PLA gene in the target wheat genome into the target Wheat realization.
  • the substance for knocking out the PLA gene in the target wheat genome may be a CRISPR/Cas9 vector.
  • the CRISPR/Cas9 vector is a CRISPR/Cas9-1 vector, which is a DNA sequence encoding a sgRNA target site designed for the PLA-A gene and the PLA-D gene (sequence 1 Positions 905-923) and the coding DNA sequence of the sgRNA target site designed for PLA-D gene (positions 939-957 of sequence 7) are inserted into pBUN411 vector together.
  • the CRISPR/Cas9 vector is a CRISPR/Cas9-2 vector, which is a sgRNA target site designed for PLA-A gene, PLA-B gene and PLA-D gene
  • the vector obtained by inserting the coding DNA sequence (sequence 1 at positions 699-718) into the pBUN411 vector.
  • the wheat of interest may be wild-type wheat material CB037.
  • the present invention provides a method for preparing wheat female parent haploid.
  • the method for preparing wheat female parent haploid includes the following steps: selfing the wheat female parent haploid induction line or its progeny prepared by the above method or crossing it as a male parent with other wheat materials to obtain selfed progeny Or the hybrid offspring is the haploid of the wheat female parent.
  • the method further includes the steps of: performing haploid trait identification and/or leaf ploidy identification and/or molecular identification on the selfed progeny or the hybrid progeny single plant, and selecting at least one method for identification as The progeny of the haploid plant is the female parent haploid of wheat.
  • the method for identifying haploid traits can be carried out according to the following method: if the plant to be tested has the characteristics of short plant, narrow leaves, overshoot, compact plant type, and male sterility, then the plant is or candidate It is a haploid; if the plant to be tested has the characteristics of plant height, wide leaves, scattered, and normal fertility, then the plant is or candidate is diploid.
  • the leaf ploidy identification method can be carried out as follows: extract the nucleus of the young leaves of the plant to be tested, and use diploid wheat leaves as a control; then use flow cytometry to detect the signal, first detect the diploid nucleus signal, and The nuclear signal peak position of diploid cells is set to 100 (since the genetic material in diploid cells is twice that of haploid cells, the haploid nuclear signal peak position appears near 50). If the nuclear signal peak of the tested plant appears near 50, the plant is or candidate is haploid; if the signal peak of the tested plant appears near 100, which is the same as the diploid nuclear signal intensity enrichment position, then the The plant is or candidate is diploid.
  • the molecular marker identification can be carried out according to the following method: PCR amplification is carried out with polymorphic primers between the male parent (maternal haploid induction line) and the female parent, and the plant to be tested is judged whether it is haploid or haploid based on the PCR amplification product Diploid: If the amplified product of the plant to be tested only has the band pattern of the maternal parent, and there is no band pattern of the male parent, the plant is or candidate is haploid; if the amplified product of the tested plant has the male parent If the plant is heterozygous with the female parent, the plant is or candidate is diploid.
  • the wheat female parent haploid induction line or the wheat female parent haploid prepared by the above method also belong to the protection scope of the present invention.
  • the present invention also provides a protein.
  • the protein provided by the present invention is a protein shown in the following a) or b) or c) or d):
  • amino acid sequence is the protein shown in sequence 3 or sequence 6 or sequence 9;
  • the present invention also provides biological materials related to the above protein.
  • the biological material provided by the present invention is any one of the following A1) to A12):
  • A1 A nucleic acid molecule encoding the above-mentioned protein
  • A2 An expression cassette containing the nucleic acid molecule described in A1);
  • A3 A recombinant vector containing the nucleic acid molecule described in A1);
  • A4 A recombinant vector containing the expression cassette described in A2);
  • A5 A recombinant microorganism containing the nucleic acid molecule described in A1);
  • A6 A recombinant microorganism containing the expression cassette described in A2);
  • A7 A recombinant microorganism containing the recombinant vector described in A3);
  • A10 A transgenic plant cell line containing the expression cassette of A2);
  • a transgenic plant cell line containing the recombinant vector described in A4) A transgenic plant cell line containing the recombinant vector described in A4).
  • nucleic acid molecule in A1) is the gene shown in 1) or 2) or 3) as follows:
  • Its coding sequence is a cDNA molecule or genomic DNA molecule shown in sequence 1 or sequence 2 or sequence 4 or sequence 5 or sequence 7 or sequence 8;
  • a cDNA molecule or genomic DNA molecule that hybridizes to the nucleotide sequence defined in 1) or 2) under stringent conditions and encodes the above-mentioned protein.
  • the substance for silencing or inhibiting the expression and/or activity of the PLA gene in the target wheat genome or knocking out the PLA gene may be a CRISPR/Cas9 vector for knocking out the PLA gene.
  • the CRISPR/Cas9 vector is the aforementioned CRISPR/Cas9-1 vector.
  • the CRISPR/Cas9 vector is the aforementioned CRISPR/Cas9-2 vector.
  • Figure 1 is a schematic diagram of the PLA gene structure and the setting of the target site using CRISPR/Cas9 technology.
  • Figure 2 shows the results of phenotypic identification.
  • the left side is diploid; the right side is haploid.
  • Figure 3 shows the results of leaf ploidy identification.
  • the left side is diploid; the right side is haploid.
  • Figure 4 shows the results of molecular identification.
  • F is the male parent
  • CS is China Spring
  • F1 is the first generation of hybrids
  • H is haploid.
  • Example 1 Obtaining wheat haploid induction genes
  • the maize haploid inducing gene ZmPLA was determined.
  • the genomic sequence of the PLA-A gene is shown in sequence 1
  • the CDS sequence is shown in sequence 2
  • the amino acid sequence of the protein encoded by the PLA-A gene is shown in sequence 3.
  • the genomic sequence of the PLA-B gene is shown in sequence 4
  • the CDS sequence is shown in sequence 5
  • the amino acid sequence of the protein encoded by the PLA-B gene is shown in sequence 6.
  • the genomic sequence of the PLA-D gene is shown in sequence 7
  • the CDS sequence is shown in sequence 8
  • the amino acid sequence of the protein encoded by the PLA-D gene is shown in sequence 9.
  • Example 2 Method for inducing the haploid of wheat female parent
  • This example uses the CRISPR/Cas9 system to knock out the PLA-A gene in the wheat A genome and/or the PLA-B gene in the B genome and/or the PLA-D gene in the D genome to obtain the following PLA gene mutations and be able to induce haploids Wheat mutants: knock out the PLA-A gene in the A genome, the PLA-B gene in the B genome and the PLA-D gene in the D genome at the same time; simultaneously knock out the PLA-A gene and the D genome in the A genome Wheat mutants with PLA-D gene; wheat mutants with the PLA-A gene in the A genome knocked out alone; wheat mutants with the PLA-A gene in the D genome knocked out alone.
  • the specific preparation method of the mutant is as follows:
  • Figure 1 is a schematic diagram of gene structure and target sites.
  • the target site sequence designed for the PLA-A gene and the PLA-D gene to knock out the two genes at the same time is CCAGGGACGTCAACCGCTT (the target site sequence is located at positions 905-923 of sequence 1 or 905-923 of sequence 7) .
  • the sgRNA target site sequence designed for the target site sequence is CCAGGGACGUCAACCGCUU, and the coding DNA sequence of the sgRNA target site is CCAGGGACGTCAACCGCTT.
  • the target site sequence for the PLA-D gene is CCCCTACATCTTCCCGCAA (the target site sequence is located at positions 939-957 in sequence 7).
  • the sgRNA target site sequence designed for the target site sequence is CCCCUACAUCUUCCCGCAA, and the coding DNA sequence of the sgRNA target site is CCCCTACATCTTCCCGCAA.
  • the target site sequence designed for the PLA-A gene, PLA-B gene and PLA-D gene to knock out the three genes at the same time is GACGGTGCTGACCATCGACG (the target site sequence is located at the 699-718th of the sequence 1 or the sequence 4 702-721 bits or 699-718 bits in sequence 7).
  • the sgRNA target site sequence designed for the target site sequence is GACGGUGCUGACCAUCGACG, and the coding DNA sequence of the sgRNA target site is GACGGTGCTGACCATCGACG.
  • the CRISPR/Cas9-1 vector is the coding DNA sequence of the sgRNA target site designed for the PLA-A gene and PLA-D gene in step 1 a and the sgRNA target site designed for the PLA-D gene in step 1 b
  • the coding DNA sequences of the two are inserted into the pBUN411 vector (pBUN411 is recorded in the following documents: Xing H L, Dong L, Wang Z P, et al. A CRISPR/Cas9 toolkit for multiplex genome editing in plants[J]. BMC plant biology, 2014 ,14(1):1.).
  • the CRISPR/Cas9-2 vector inserts the coding DNA sequences of the sgRNA target sites designed for the PLA-A gene, PLA-B gene and PLA-D gene in step 1 c into the pBUN411 vector (pBUN411 is described in the following document: Xing H L, Dong L, Wang Z P, et al. A CRISPR/Cas9 toolkit for multiplex genome editing in plants[J]. BMC plant biology, 2014, 14(1):1.).
  • CRISPR/Cas9-1 vector and CRISPR/Cas9-2 vector were transformed into Agrobacterium competent cell EHA105 (purchased from Huayueyang Biotechnology Co., Ltd., publicly available through purchase) through heat shock transformation, respectively, and recombinant bacteria were obtained.
  • T0 generation transgenic wheat plants were obtained after screening, differentiation and rooting according to conventional methods.
  • T0 generation transgenic wheat plants were collected, and genomic DNA was extracted as a template, and PCR amplification was performed with the following primers to obtain PCR amplification products of different strains.
  • PLA-A 4AF: GTCAAGATCTCCAGCCGAGAC;
  • PLA-B 4BF: AACTCAACATGGGGCGTCCTC;
  • PLA-D 4DF: TTCGGGTCCGGATTCTATTGTG;
  • the PCR products of different strains were sequenced by Sanger and compared with the wild-type wheat PLA gene according to the sequencing results to identify whether the PLA-A gene, PLA-B gene and PLA-D gene in different strains of T0 generation transgenic wheat Mutation occurred. Plants with mutations in the PLA gene were recorded as positive T0 generation transgenic wheat.
  • the positive T0 generation transgenic wheat obtained in the above step 4 is harvested and then sown to obtain T1 generation transgenic wheat.
  • the specifics are as follows: using the genomic DNA of the T1 generation transgenic wheat as a template, using the PLA mutation sequence detection primers for amplification, the PCR product is subjected to Sanger sequencing, and the T1 generation transgenic according to the sequencing results The mutation of PLA gene in wheat is described.
  • the PLA-A genes on the two homologous chromosomes in the A genome of the T1 transgenic wheat PLA gene mutant strain Da14-1 are both PLA-A-1 mutant genes, and the PLA on the two homologous chromosomes in the D genome -D genes are all PLA-D-1 mutant genes.
  • the PLA-A-1 mutant gene is the gene sequence obtained by deleting the 909th base G of the PLA-A gene shown in sequence 1; the PLA-D-1 mutant gene is the PLA-D gene shown in sequence 7 The gene sequence obtained after deletion of GG at positions 908-909.
  • the PLA-A genes on the two homologous chromosomes in the A genome of the T1 transgenic wheat PLA gene mutant strain Da14-1-A are both PLA-A-1 mutant genes.
  • the PLA-A-1 mutant gene is a gene sequence obtained by deleting the 909th base G of the PLA-A gene shown in SEQ ID NO:1.
  • the PLA-D genes on the two homologous chromosomes in the D genome of the T1 generation transgenic wheat PLA gene mutant strain Da14-1-D are both PLA-D-1 mutant genes.
  • the PLA-D-1 mutant gene is a gene sequence obtained by deleting the GG at positions 908-909 of the PLA-D gene shown in SEQ ID NO: 7.
  • the PLA-A gene on the two homologous chromosomes in the A genome of the T1 transgenic wheat PLA gene mutant line Ne147-1 are both PLA-A-2 mutant genes, and the PLA on the two homologous chromosomes in the B genome -B genes are both PLA-B-1 mutant genes, and the PLA-D genes on the two homologous chromosomes in the D genome are both PLA-D-2 mutant genes.
  • the PLA-A-2 mutant gene is the gene sequence obtained by deleting the TCGA at bases 713-716 of the PLA-A gene shown in sequence 1; the PLA-B-1 mutant gene is the PLA- shown in sequence 4.
  • T1 generation transgenic wheat PLA gene mutant lines (T1 generation transgenic wheat PLA gene mutant line Da14-1, T1 generation transgenic wheat PLA gene mutant line Da14-1-A, T1 generation transgenic wheat PLA gene mutant line Da14- 1-D, T1 generation transgenic wheat PLA gene mutant line Ne147-1) materials were selfed to obtain selfed progeny.
  • T1 generation transgenic wheat PLA gene mutant lines (T1 generation transgenic wheat PLA gene mutant line Da14-1, T1 generation transgenic wheat PLA gene mutant line Da14-1-A, T1 generation transgenic wheat PLA gene mutant line Da14- The pollen of the 1-D and T1 transgenic wheat PLA gene mutant strain Ne147-1) was granted to wild-type wheat material, China Spring, to be crossed to obtain hybrid offspring. At the same time, the pollen of wild-type wheat CB037 (with no mutation in the PLA gene) was granted to the offspring of wild-type wheat China Spring as a control.
  • the progeny obtained above were sown in the field, and the phenotype of the individual plants of the progeny was observed.
  • the haploid has the characteristics of short plants, narrow leaves, and upswing, compact plant type, and male sterility.
  • the diploid is characterized by tall plants and leaves. It is wide, loose and fertile ( Figure 2).
  • T1 generation transgenic wheat PLA gene mutant line Da14-1 selfed 154 progeny, 17 out of 154 progeny showing haploid traits single plant, proposed to be haploid plants;
  • One of the 225 progenies of the T1 generation transgenic wheat PLA gene mutant line Da14-1-D self-bred was obtained as a single plant with haploid traits, which was proposed as a haploid plant.
  • the haploid plants in the hybrid offspring of the mutant material in the above step 1 and the Chinese spring were used for genotype identification using polymorphic molecular markers (Xbarc284 and xgwm124-1B). The results are shown in Figure 4, and the results show that the haploid plants only have the band type of Chinese Spring (mother parent). It is further proved that the haploid obtained by this method is parthenogenetic maternal haploid.
  • the primer sequence of the polymorphism marker is as follows:
  • Xbarc284-L GCGTCAGAAATGCAAGAAAAATAGG;
  • Xbarc284-R GCGGAAGAAAAGGACGAAGACAAG;
  • Xgwm124-1B-F GCCATGGCTATCACCCAG;
  • Xgwm124-1B-R ACTGTTCGGTGCAATTTGAG.
  • the T1 generation transgenic wheat PLA gene mutant line is selfed or crossed with hybrids, if it is identified as haploid according to any of the above three methods, the plant is or candidate It is a wheat female parent haploid; if none of the above three methods is a haploid, the plant is not or the candidate is not a wheat female parent haploid.
  • the statistical results are shown in Table 1: It can be seen that the T1 generation transgenic wheat PLA gene mutant line Ne147-1 has the highest haploid induction rate, and the T1 generation transgenic wheat PLA gene mutant strain It is followed by Da14-1.
  • the PLA gene mutant plants obtained by mutating the PLA-A gene in the wheat A genome and/or the PLA-B gene in the wheat B genome and/or the PLA-D gene in the wheat D genome can be used as wheat A female parent haploid induction line, which is selfed or crossed with other wheat materials to obtain a wheat female parent haploid in the offspring.
  • the present invention obtains the homologous gene PLA encoding wheat phospholipase by analyzing the homologous genes of the induced gene ZmPLA1 in maize in wheat.
  • the gene exists in the three chromosomes A, B, and D of wheat, respectively named PLA -A gene, PLA-B gene and PLA-D gene, through CRISPR/Cas9 site-directed mutagenesis technology and transgenic experiments, successfully obtained PLA gene mutation transgenic materials, using transgenic materials for selfing or hybridization with other materials, in the offspring A certain proportion of haploid plants was observed, which verified that the wheat material after PLA mutation has the function of inducing the haploid of the female parent of wheat.
  • the present invention not only lays an important foundation for revealing the genetics and biological mechanism of wheat female parent haploid production, but also is important for breeding new induction lines, further increasing the induction rate, and improving the efficiency of wheat haploid species. Meaning.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Microbiology (AREA)
  • Biophysics (AREA)
  • Cell Biology (AREA)
  • Physics & Mathematics (AREA)
  • Plant Pathology (AREA)
  • Developmental Biology & Embryology (AREA)
  • Botany (AREA)
  • Environmental Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Virology (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)

Abstract

Gène haploïde induit de blé et application associée. Un gène de PLA codant pour la phospholipase de blé est obtenu par analyse du gène homologue du gène induit ZmPLA1 du maïs dans le blé, qui est présent dans les trois groupes de chromosomes du blé A, B et D et appelés respectivement PLA-A, PLA-B Et PLA-D. Un matériel transgénique ayant une mutation de gène de PLA est obtenu par la technologie de mutagenèse et l'expérience transgénique dirigées sur site , et l'utilisation du matériel transgénique pour l'autogamie ou une hybridation avec d'autres matériels, une certaine proportion de plantes haploïdes sont observées dans la descendance, le matériel de blé après la mutation du PLA est vérifié afin qu'il présente la fonction d'induire la production d'haploïde maternel de blé.
PCT/CN2020/073688 2019-01-31 2020-01-22 Gène haploïde induit de blé et application associée WO2020156406A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201910098036 2019-01-31
CN201910098036.0 2019-01-31

Publications (1)

Publication Number Publication Date
WO2020156406A1 true WO2020156406A1 (fr) 2020-08-06

Family

ID=70324121

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2020/073688 WO2020156406A1 (fr) 2019-01-31 2020-01-22 Gène haploïde induit de blé et application associée

Country Status (2)

Country Link
CN (1) CN111073875B (fr)
WO (1) WO2020156406A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116240219A (zh) * 2023-02-15 2023-06-09 青岛农业大学 一种小麦rth-1基因及其应用

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114957422B (zh) * 2022-06-15 2024-08-27 中国水稻研究所 一种诱导单倍体的方法及其在植物育种中的应用
CN117069814B (zh) * 2020-12-03 2024-04-19 中国农业大学 孤雌生殖单倍体诱导基因GhDMP及其应用

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0584117A1 (fr) * 1991-04-26 1994-03-02 Zeneca Limited Modification de la synthese de la lignine dans les plantes
CN106701803A (zh) * 2017-01-13 2017-05-24 中国农业大学 玉米母本单倍体主效诱导基因及应用
US20180092316A1 (en) * 2013-03-15 2018-04-05 Syngenta Participations Ag Haploid induction compositions and methods for use therefor
CN108192912A (zh) * 2017-01-13 2018-06-22 中国农业大学 一种诱导产生玉米母本单倍体的方法
CN109072178A (zh) * 2015-11-18 2018-12-21 先正达参股股份有限公司 单倍体诱导组合物及其使用方法

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0584117A1 (fr) * 1991-04-26 1994-03-02 Zeneca Limited Modification de la synthese de la lignine dans les plantes
US20180092316A1 (en) * 2013-03-15 2018-04-05 Syngenta Participations Ag Haploid induction compositions and methods for use therefor
CN109072178A (zh) * 2015-11-18 2018-12-21 先正达参股股份有限公司 单倍体诱导组合物及其使用方法
CN106701803A (zh) * 2017-01-13 2017-05-24 中国农业大学 玉米母本单倍体主效诱导基因及应用
CN108192912A (zh) * 2017-01-13 2018-06-22 中国农业大学 一种诱导产生玉米母本单倍体的方法

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
DATABASE GENBANK 24 February 2017 (2017-02-24), "patatin-like protein 1 [Aegilops tauschii subsp. tauschii", XP055729110, Database accession no. XP_020152811 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116240219A (zh) * 2023-02-15 2023-06-09 青岛农业大学 一种小麦rth-1基因及其应用
CN116240219B (zh) * 2023-02-15 2024-03-29 青岛农业大学 一种小麦rth-1基因及其应用

Also Published As

Publication number Publication date
CN111073875A (zh) 2020-04-28
CN111073875B (zh) 2022-01-25

Similar Documents

Publication Publication Date Title
CN109628480B (zh) 玉米孤雌生殖单倍体诱导基因ZmPLA1E及其应用
CN111996209B (zh) 孤雌生殖单倍体诱导基因dmp及其应用
CN111763687B (zh) 一种基于基因编辑技术快速培育玉米单倍体诱导系的方法
CN108192912B (zh) 一种诱导产生玉米母本单倍体的方法
CN106701803B (zh) 玉米母本单倍体主效诱导基因及应用
WO2020156406A1 (fr) Gène haploïde induit de blé et application associée
WO2019000806A1 (fr) Procédé de création d'une lignée mâle stérile de solanum lycopersicum par édition du génome, et application associée
WO2018129704A1 (fr) Gène induisant un effet majeur d'haploïde parent femelle de maïs et application
CN109554387A (zh) 一种诱导产生水稻母本单倍体的方法
CN103589805B (zh) 赋予玉米斐济病毒抗性的主要qtls
US20220333125A1 (en) GENE ZmPLD3 FOR INDUCING MAIZE MATERNAL HAPLOID PRODUCTION AND ITS APPLICATION THEREOF
CN108794610B (zh) 玉米杂交不亲和相关蛋白ZmGa1S及其编码基因与应用
WO2022117024A1 (fr) Gène d'induction haploïde parthénogénétique et son application
CN112195269B (zh) 与水稻核雄性不育表型相关的分子标记和应用
CN111100869B (zh) 一种与水稻光温敏核雄性不育性状共分离的分子标记和应用
CN112795576A (zh) 一种谷子单倍体诱导基因SiMTL及其应用
CN114686613A (zh) 杂交水稻高产优质基因GW3p6的分子标记及其应用
CN115466748B (zh) 诱导玉米单倍体产生的基因ZmKNL2及应用
MX2012004639A (es) Medios para inducir la apomixis en plantas cultivadas para reproduccion sexual y el uso para la produccion de plantas total o parcialmente apomicticas.
CN116376934A (zh) NLR基因对Pm55/SuPm55及其抗小麦白粉病育种利用
CN117512170A (zh) 水稻雄性不育基因OsGSL2的分子标记及其应用
CN118638956A (zh) 与辣椒细胞核雄性不育基因连锁的分子标记、基因分型引物和应用
CN115976055A (zh) 一种玉米矮秆基因及其分子标记
CN116063423A (zh) 一种提高单倍体诱导效率的突变型OsPLA1m1及其应用
CN111690047A (zh) 一个玉米细胞核雄性育性基因ipe2的克隆与应用

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 20747683

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 20747683

Country of ref document: EP

Kind code of ref document: A1