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Method for creating apple non-fusion allopolyploid rootstock based on whole genome mutagenesis

An allopolyploid and whole-genome technology, applied in the field of creating apple apoplexy allopolyploid rootstocks based on whole-genome mutagenesis, can solve problems such as difficulty in creation, restrictions on the application of apomixis, and high degree of apomixis, and achieve Save time and space for screening, easy to master operating techniques, and broad application prospects

Active Publication Date: 2022-03-25
SHANDONG AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, there are currently difficulties in creating seedling rootstocks with apomictic characteristics, which restricts the application of apomictic reproduction in breeding programs and asexual seed production
Especially in the breeding of apple rootstocks, there are problems such as long childhood period, large plant space, long screening cycle, and high apomixis traits cannot be passed on to offspring through sexual crossing

Method used

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  • Method for creating apple non-fusion allopolyploid rootstock based on whole genome mutagenesis
  • Method for creating apple non-fusion allopolyploid rootstock based on whole genome mutagenesis
  • Method for creating apple non-fusion allopolyploid rootstock based on whole genome mutagenesis

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Embodiment 1

[0037] A method for creating apple afused allopolyploid rootstock based on whole-genome mutagenesis of the present invention comprises the following steps:

[0038] (1) Whole-genome sequencing of apple apomictic rootstocks: The whole-genome sequencing of apple apomictic rootstocks is performed by, but not limited to, PacBio Hifi sequencing, Oxford Nanopore Technology sequencing, and Illumina next-generation sequencing platforms. The obtained raw data were assembled, the assembly results were corrected with NextPolish software and second-generation Illumina data, and high-quality genomes were assembled. Using published apple genome sequences such as Hanfu, Jinguan, etc. and Hi-C data, ALLHiC software was used for Contig analysis. Phase separation and mounting, and finally obtain the genome sequence of the apomixis target material mounted on all chromosomes, perform Gap Closing on the genome mounted on the chromosome using the ONT sequencing data corrected by NECAT, and then perf...

Embodiment 2

[0047] Now take Pingyi sweet tea as an example, the method for creating apple afused allopolyploid rootstock based on whole genome mutagenesis according to the present invention is introduced in detail, and the method comprises the following steps:

[0048] (1) After sampling the young leaves of a single 100-year-old wild 'Pingyi Sweet Tea' tree in Yimeng Mountains, they were quickly frozen with liquid nitrogen, and sent to Beijing Baimaike Biotechnology Co., Ltd. for sequencing, and the genome extraction was completed by Baimaike Company;

[0049] (2) Whole genome sequencing was performed using PacBio Hifi sequencing, Oxford Nanopore Technology sequencing and Illumina second-generation sequencing platform, and the PacBio Hifi data was assembled using Canu hifi software. The assembly results were corrected with NextPolish software and second-generation Illumina data. Rich genome sequence and Hi-C data, ALLHiC software was used to separate and mount Contig, and finally the genom...

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Abstract

The invention discloses a method for creating apple apomixis allopolyploid rootstocks based on whole genome mutagenesis. The method comprises the following steps: performing whole genome sequencing on apple apomixis rootstocks; after removing stigmas of the single plants of the sequencing woody material, harvesting seeds; the harvested seeds are subjected to low-temperature refrigeration and lamination; mutagenizing EMS with different concentrations, germinating seeds and sowing; collecting the seeds for laminated germination; performing water culture and identifying phenotypes; carrying out differential material plant tissue culture, and inducing rooting to identify a root phenotype; performing whole genome re-sequencing and transcriptome sequencing on the differential material; finding a mutagenic gene; and carrying out mutagenic gene function verification on an agrobacterium rhizogenes transformation system. According to the method, the novel non-fusion rootstocks are rapidly screened and identified through apple apomixis characteristics, root phenotype analysis and transcriptome high throughput, the screening time and space are greatly saved, the method process is simple, the operation technology is easy to master, and the method has wide application prospects.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture, and in particular relates to a method for creating apple afused allopolyploid rootstocks based on whole genome mutagenesis. Background technique [0002] Apomixis refers to a special asexual reproduction that directly produces asexual embryos and asexual seeds without the fertilization and combination of sperm and egg cells. The offspring do not produce segregation, thus exhibiting a high degree of uniformity and other excellent traits, which can be preserved during reproduction. Compared with the existing sexual seeds as base stock and clonal dwarf stock used in apple cultivation, apomictic rootstocks have many advantages: through asexual seed propagation, the offspring are neat and consistent; the reproduction coefficient is high, compared with cuttings, layering and other reproduction methods. , high reproductive efficiency, strong root system, good site performance, strong adapt...

Claims

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Application Information

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IPC IPC(8): A01G17/00A01G31/00A01G2/10C12Q1/6895C12N15/29C07K14/415C12N15/82A01H5/06A01H6/74
CPCA01G17/005A01G31/00A01G2/10C12Q1/6895C07K14/415C12N15/8205C12N15/8261A01H5/06A01H6/74C12Q2600/13C12Q2600/156
Inventor 张世忠张步航郑成超李菡齐琦姜舒纳
Owner SHANDONG AGRICULTURAL UNIVERSITY
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