Wheat-diploid agropyron elongatum 45K liquid chip and application

A liquid-phase chip and wheatgrass technology, which is applied to DNA/RNA fragments, biochemical equipment and methods, and the determination/inspection of microorganisms, etc., can solve the problems of ineffective identification, the development of liquid-phase chips for wheat-long-eared wheatgrass, etc. , to achieve the effect of reducing detection cost, high detection accuracy, and more effective information

Active Publication Date: 2022-05-10
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, single nucleotide liquid-phase chips based on GBTS technology have been developed and utilized in barley, cotton, corn, rice, soybean and other plants, but there is no liquid-phase chip developed for wheat-Thinopyrum elongatum, so it is impossible to Efficient identification and analysis of materials formed by distant hybridization of wheat and Thiopyrum elongatum

Method used

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  • Wheat-diploid agropyron elongatum 45K liquid chip and application
  • Wheat-diploid agropyron elongatum 45K liquid chip and application
  • Wheat-diploid agropyron elongatum 45K liquid chip and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] A wheat-diploid E. elongatum 45K liquid-phase chip contains a set of wheat genome-wide 5K oligonucleotide probe sites and a set of diploid E. elongatum genome-wide 40K oligonucleotide probe sites Dots, ie, predominantly 40K E. elongatum oligonucleotide sites supplemented by 5K wheat oligonucleotide sites.

[0029] Specifically, the liquid phase chip is developed based on targeted capture technology. Its working principle is based on designing probes and detecting sample DNA, and performing targeted capture and sequencing through complementary base pairing, so as to realize gene detection and typing in target regions. the goal of.

[0030] The specific method for screening 5K oligonucleotide sites in the whole wheat genome is as follows.

[0031] (1) Download the resequenced sequence of diploid Echinopsis elongatum (NCBI database accession number SRR10023775), and use BWA software to compare the sequence with the Chinese spring wheat reference genome (Ensemble Genome da...

Embodiment 2

[0044] A wheat-diploid E. elongatum 45K liquid-phase chip, including a set of 5K oligonucleotide probe sites for the whole genome of wheat and a set of 40K oligonucleotide probes for the whole genome of diploid E. elongatum location.

[0045] Among them, the genome distribution of the core sites of the 5K oligonucleotide probe sites in the whole wheat genome is as follows: figure 1 As shown, at the same time, its core site information is shown in Table 1.

[0046] Table 1. Core site information of 5K oligonucleotide probe sites in the whole wheat genome

[0047]

[0048]

[0049]

[0050]

[0051] In the locus numbers in Table 1, 1A~7A, 1B~7B, and 1D~7D on the left indicate the chromosomes where the loci are located, and the subsequent two sets of values ​​are the oligonucleotide probes on the wheat reference genome (Chinese spring). Each chromosome corresponds to the start and end sites; the whole genome sequence version number of the wheat reference genome Chin...

Embodiment 3

[0065] This example is the identification of the diploid Etyophony elongatum and wheat varieties Zhongguochun and Aikang 58 by the liquid-phase chip.

[0066] (1) DNA extraction and quality inspection of the material to be tested: select diploid Echinopsis elongatum, wheat variety China Chunhe Aikang 58, take fresh tissue of the sample to be tested, and use the CTAB method to extract DNA; use agarose gel electrophoresis Sample DNA was tested to ensure the integrity of the genome; the concentration of genomic DNA was measured using a micro-volume ultraviolet spectrophotometer, and the working concentration of DNA in each sample was adjusted to 10-50ng / ul. Among them, the wheat variety China Spring included two biological replicates.

[0067] (2) 45K liquid-phase chip detection of wheat-diploid Etypyronium elongatum: targeted gene capture technology based on liquid-phase probe hybridization (GenoBaits: http: / / www.molbreeding.com / index.php / Technology / GenoBaits .html) The standard...

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Abstract

The invention belongs to the technical field of whole genome gene chips, and relates to a wheat-diploid agropyron elongatum 45K liquid chip and application thereof. The liquid phase chip comprises a group of wheat whole genome 5K oligonucleotide probe sites and a group of diploid agropyron elongatum whole genome 40K oligonucleotide probe sites. According to the liquid-phase chip, 5K wheat and 40K diploid agropyron elongatum oligonucleotide sites are integrated into the same chip, the composition and change of exogenous and wheat genetic materials in the distant hybridization process can be monitored only through one-time detection, and the liquid-phase chip is high in detection accuracy, more in effective information, high in flexibility and diversity and low in detection cost; important technical support is provided for wheat molecular breeding.

Description

technical field [0001] The invention belongs to the technical field of whole-genome gene chips, and relates to a wheat-diploid Etynium elongatum 45K liquid phase chip and its application, in particular to a wheat-diploid Etynium elongatum obtained based on targeted capture sequencing technology 45K liquid phase chip and its application. Background technique [0002] Wheat is an important food crop in my country, and its high and stable yield is crucial to food security. Broadening the genetic diversity of wheat breeding parents can lay the foundation for breaking through the bottleneck of yield and improving quality, stress resistance and disease resistance. Among them, the development and utilization of wheat relatives based on distant hybridization is one of the most important means. Through the long-term practice and efforts of many scientific researchers, at present, people have realized the distant hybridization of more than 20 close relatives of wheat and wheat. Elo...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/6895C12Q1/6837
CPCC12Q1/6895C12Q1/6837C12Q2600/156
Inventor 吉万全邓平川杜欣王斯文程小方陈春环赵继新王长有李停栋
Owner NORTHWEST A & F UNIV
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