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Method for conducting high-throughput-testing on wheat gibberellic-disease-resisting genetic types and kit thereof

A wheat scab, high-throughput technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve problems such as wheat quality and production loss

Active Publication Date: 2018-05-08
CHINA GOLDEN MARKER BEIJING BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Wheat head blight is mainly caused by Fusarium graminearum (Fusarium graminearum) and yellow Fusarium (Fusarium culmorum). Injured, causing great loss to wheat quality and production

Method used

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  • Method for conducting high-throughput-testing on wheat gibberellic-disease-resisting genetic types and kit thereof
  • Method for conducting high-throughput-testing on wheat gibberellic-disease-resisting genetic types and kit thereof
  • Method for conducting high-throughput-testing on wheat gibberellic-disease-resisting genetic types and kit thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0109] The selection of the SNP molecular marker of embodiment 1 wheat scab resistance gene (FHB1 and FHB5) locus

[0110] The FHB1 gene is located on wheat chromosome 3B, and the FHB5 gene is located on wheat chromosome 5A (refer to wheat Chinese spring genome Refseqv1.0 version), by comparing the Chinese spring gene genome (Genbank No.: FN564434.1 and HE774676.1) 3 Genome (Genbank No.: KX907434.1) sequence, obtained 3 SNP sites on chromosome 3B, selected Zhongyujin marked wheat 600K SNP chip, selected 3 SNP sites in the interval 107,277,266 and 109,306,268, a total of 6 The SNP sites were used for marker development, see Table 1 below.

[0111] Table 1 Information of SNP sites linked to FHB1 and FHB5

[0112] (The chromosomal position is the position of the Chinese spring V1 version of the wheat reference genome)

[0113]

[0114]

[0115] According to the commercially available KASP Master Mix detection kit from LGC Company, Primer3 primer design software was used t...

Embodiment 2

[0121] Example 2 High-throughput detection of SNP molecular markers at the wheat head blight resistance gene FHB1 locus and FHB5 locus

[0122] The 6 KASP primer sets designed in Example 1 were used, and wheat genomic DNA was used as a template to analyze the SNP sites after PCR amplification.

[0123] 1. Extract the sample genomic DNA:

[0124] According to the plant genomic DNA extraction kit (catalogue number: DP-305) provided by TIANGEN Company, and extract the genomic DNA of the wheat sample for testing according to the kit instructions, the specific steps are as follows:

[0125] (1) Grinding 100 mg of fresh or 20°C frozen sample material in liquid nitrogen;

[0126] (2) Quickly transfer the ground powder to a centrifuge tube pre-installed with 700 μl of buffer solution GP1 preheated at 65°C. The buffer solution GP1 contains mercaptoethanol with a final concentration of 0.1wt%, and quickly mix it upside down Finally, place the centrifuge tube in a 65°C water bath for 2...

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Abstract

The invention relates to a method for conducting high-throughput-testing on wheat gibberellic-disease-resisting genetic types and a kit thereof, in particular to SNP molecular markers used for conducting high-throughput-testing on wheat gibberellic-disease-resisting genes, an identifying method thereof, a method used for detecting the SNP molecular markers, a primer set and the kit.

Description

technical field [0001] The invention relates to molecular identification of wheat scab resistance, in particular to high-throughput detection of molecular markers used for molecular identification of wheat scab resistance. Background technique [0002] Wheat head blight (Fusarium head blight, FHB) is one of the main diseases of wheat. Wheat head blight is mainly caused by Fusarium graminearum (Fusarium graminearum) and yellow Fusarium (Fusarium culmorum). damage to wheat quality and production. The most economical and effective way to control wheat scab is to adopt advanced molecular breeding methods, make full use of the resistance resources in the wheat resource bank, and carry out disease resistance breeding and disease resistance improvement. [0003] Therefore, it is urgent to develop methods and kits suitable for high-throughput detection of molecular markers related to wheat scab resistance, as well as capable of high-throughput detection of molecular markers relate...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 翟晨光娄璐岩李梦安霞柴宇超高玉峰卢洪
Owner CHINA GOLDEN MARKER BEIJING BIOTECH CO LTD
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