The related gene tadir-b1 of wheat stem rot and its application

A technology of stalk rot and tadir-b1, applied in application, genetic engineering, plant genetic improvement, etc., can solve the problems of differences in resistance of wheat varieties, inconsistent identification conditions and evaluation standards, and slow progress in gene mining. To achieve the effect of saving time and resources

Active Publication Date: 2022-07-08
HENAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, due to the inconsistency of identification conditions and evaluation standards for wheat stalk rot, the differences in resistance of wheat varieties, and the limitation of complex genetic backgrounds, the mining of genes related to stalk rot in wheat has progressed slowly; so far, the control The cloning of wheat stem rot related genes is still rarely reported

Method used

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  • The related gene tadir-b1 of wheat stem rot and its application
  • The related gene tadir-b1 of wheat stem rot and its application
  • The related gene tadir-b1 of wheat stem rot and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1: Identification of stem rot resistance of different wheat varieties

[0058] 1. Test strain selection, experimental soil preparation and medium preparation

[0059] The strain used in the test is Fusarium pseudograss WZ-8A, which is currently a popular strain in the Huanghuai wheat area.

[0060] The soil used in the experiment was loam with moderate fertility. Before using the soil, weeds and large clods were removed with an 18-mesh sieve, and then sterilized in a sterilizing pot of 0.1 Mpa at 121 °C for 1 h for use.

[0061] The medium used in the experiment included PDA medium and millet medium. Wherein, the preparation method of PDA culture medium is: choose the fresh potatoes of moderate size, peel and cut into medium thin pieces, take 200g thin pieces and put them in the pot, add distilled water and boil for about 20min, when the potato pieces can be easily smashed with a glass rod Filter with four layers of gauze. The filtrate was returned to the pot...

Embodiment 2

[0066] Example 2: Wheat stem rot-related genes TaDIR-B1 locating and obtaining

[0067] Through phenotypic identification, 435 wheat varieties with different resistance to stem rot were distinguished, and further combined with population 660KSNP chip information (Beijing Compson Biotechnology Co., Ltd.), genome-wide association analysis (GWAS) was performed after quality control to identify SNP loci significantly associated with wheat stem rot, such as figure 1 shown. Haplotype analysis was performed on the identified significant SNP loci on chromosome 4B, and the results were as follows figure 2 As shown, it shows that a block is formed and contains 5 haplotypes, and the disease index of the corresponding materials of different haplotypes is significantly different.

[0068] Further utilizing the above-mentioned SNP site, to EnsemblPlants The Chinese spring data information in the database was blasted, and it was found that these SNP loci-locked segments were located ...

Embodiment 3

[0078] Embodiment three: TaDIR-B1 Allelic marker development and application

[0079] According to the G / A separation of the 15th base from ATG in the sequence of SEQ ID NO.1 between different materials, a dCAPS marker was developed to detect this mutation site, and the differential site-specific primer pair included:

[0080] The nucleotide sequence of the upstream primer sequence shown in SEQ ID NO.6 is 5'-ACTTTTGCGTCGTAGTGAGC-3' and

[0081] The nucleotide sequence of the downstream primer sequence shown in SEQ ID NO. 7 is 5'-GGAGGACGAGAAGCAGCGGCAGTGG-3'.

[0082] Utilize specific restriction endonucleases Bal1 The amplification product is digested with enzyme digestion, wherein the sequence with the differential site G in the amplification product is processed by Bal1 Wheat varieties / lines with 170 bp after digestion were named as TaDIR-B1a Allelic genotype, the sequence with the difference site A in the amplified product was Bal1 Wheat varieties / lines that are s...

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Abstract

The invention relates to a gene related to wheat stem rot TaDIR‑B1 and its applications. The gene sequence is shown in SEQ ID NO.1, and the 15th base from ATG at the 5' end is G or A, wherein type A causes the codon to change from TGG to TGA, resulting in premature termination of amino acid translation; the present invention provides wheat stem rot gene TaDIR‑B1 sequence, the protein sequence encoded by the gene, and identification TaDIR‑B1 Gene-specific primer pairs, and dCAPS markers were developed for identification TaDIR‑B1 Different allele types; the present invention also constructs a silencing vector based on the above gene, which is applied to the functional verification of the gene. The discovery, confirmation and application of the wheat stem rot-related gene of the present invention not only helps to reveal the genetic basis of the wheat stem rot, but also plays an important role in improving the quality of wheat by using genetic engineering technology.

Description

technical field [0001] The invention relates to the technical field of biological genetic engineering, in particular to a wheat stem rot-related gene TaDIR- B1 and its applications. Background technique [0002] Wheat stem rot ( crown rot , CR) is a soil-borne disease caused by a variety of pathogenic fungi, also known as dry soil foot rot ( dry land foot rot ), sickle root rot ( Fusarium root rot ) and sickle stem rot ( Fusarium crown rot )Wait. The disease is a worldwide intractable disease, and its occurrence has been reported in more than 10 countries, including the United States, Australia, Italy, Turkey, Canada, etc. ( Paulitz et al., 2002; Smiley et al., 2005; Hollaway et al., 2013). In my country, Li et al. (2012) reported for the first time that Fusarium graminearum ( Fusarium pseudograminerum ) caused by wheat stem base rot. According to the investigation by the research group of wheat soil-borne diseases from the School of Plant Protection of ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12N15/82C12N15/11C07K14/415C12Q1/6895A01H5/00A01H6/46
CPCC07K14/415C12Q1/6895C12N15/8218C12N15/8282C12Q2600/13C12Q2600/156C12Q2600/172
Inventor 陈锋阳霞赵磊任妍张宁董中东孙丛苇
Owner HENAN AGRICULTURAL UNIVERSITY
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