Method for rapidly identifying thinopyrum intermedium 4St chromosome

A technology of Thinopyrum and chromosomes, which is applied in the field of plant molecular genetics and breeding, and can solve problems such as limited recognition

Active Publication Date: 2021-08-17
SHANXI AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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  • Method for rapidly identifying thinopyrum intermedium 4St chromosome
  • Method for rapidly identifying thinopyrum intermedium 4St chromosome
  • Method for rapidly identifying thinopyrum intermedium 4St chromosome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] The detection of molecular marker sacThiC11-05 in the plant material to be tested (nucleotide sequence is:

[0041] TGAAGCCACGACTCTTACCCTACTTCAGCCATTCAATTTTGTATACCATCAATTCAGAGTCCATCATCTTCCTGATGTAATCCTAGACAACTTTTGCAAAGCTCC, such as SEQ ID NO. 1)

[0042] The primers for detecting molecular marker sacThiC11-05 are sacThiC11-05-F and sacThiC11-05-R, and their sequences are as follows:

[0043] sacThiC11-05-F: 5'-TGAAGCCACGACTCTTACCT-3', such as SEQ ID NO.7;

[0044] sacThiC11-05-R: 5'-GGAGCTTTGCAAAAGTTGTCT-3', such as SEQ ID NO.8.

[0045] The detection process is as follows:

[0046] Extract the genomic DNA of the plant to be detected, use the extracted genomic DNA as a template, and use sacThiC11-05-F and sacThiC11-05-R as primers for PCR amplification. After the amplification is completed, separate the amplified product by electrophoresis to amplify The corresponding 104bp DNA band is the plant material containing the 4St chromosome or 4St chromosome fragment of Thip...

Embodiment 2

[0053] Detection of the molecular marker sacThiC11-15 in the plant material to be tested (CTGCTCCAGCTCAGAACAATCCCTCCCTGCAGCGCACGCGGAACCAGCACGCATTTGTTCGGCGCCCGCCCCGAGGCCGGCCGTCCGGTGTCGCAATCCCGTCGTTCCCAGTCCCGTCCCCCGTCAAACGGAATCCTCGCCGGGAGGTGAACCCAACGAAACAAAC, such as SEQ ID NO.2)

[0054] The primers for detecting the molecular marker sacThiC11-15 are sacThiC11-15-F and sacThiC11-15-R, and their sequences are as follows:

[0055] sacThiC11-15-F: 5'-CTGCTCCAGCTCAGAACAAT-3', such as SEQ ID NO.9;

[0056] sacThiC11-15-R: 5'-GTTTGTTTCGTTGGGTTCAC-3', such as SEQ ID NO.10.

[0057] The detection process is as follows:

[0058] Extract the genomic DNA of the plant to be detected, use the extracted genomic DNA as a template, and use sacThiC11-15-F and sacThiC11-15-R as primers for PCR amplification. After the amplification is completed, the amplified product is separated by electrophoresis to amplify The corresponding 165bp DNA band is the plant material containing the 4St chromosome ...

Embodiment 3

[0062] Detection of the molecular marker sacThiC11-25 in the plant material to be tested (GTGGAGCTTTTGCTTCTGCCCGAGCATTTCAGAAAAAAGCAGGTGGCCTTTTGTTGCTGTAGCACATACACACACATTACAGATAGAGAGCTCGTTAGGCAGAGAGAAGGAGCCTGCGTGAACTAGAACTGCAGGGGAACCTTGAGCAACGAATA, such as SEQ ID NO.3)

[0063] The primers for detecting the molecular marker sacThiC11-25 are sacThiC11-25-F and sacThiC11-25-R, and their sequences are as follows:

[0064] sacThiC11-25-F: 5'-GTGGAGCTTTTGCTTCTGC-3', such as SEQ ID NO.11;

[0065] sacThiC11-25-R: 5'-TATTCGTTGCTCAAGGTTCC-3', such as SEQ ID NO.12.

[0066] The detection process is as follows:

[0067] Extract the genomic DNA of the plant to be detected, use the extracted genomic DNA as a template, and use sacThiC11-25-F and sacThiC11-25-R as primers for PCR amplification. After the amplification is completed, the amplified product is separated by electrophoresis to amplify The corresponding 155bp DNA band is the plant material containing the 4St chromosome or 4St chro...

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Abstract

The invention discloses a method for rapidly identifying an thinopyrum intermedium 4St chromosome, and relates to the technical field of plant molecular genetic breeding. The method comprises the following steps: plant genome DNA is extracted, and PCR amplification is carried out; and if any target fragment is obtained through amplification, an thinopyrum intermedium 4St chromosome gene exists. The molecular marker disclosed by the invention can be used for rapidly detecting the 4St chromosome or the 4St chromosome segment in chromosome engineering lines (chromosome addition lines, substitution lines, translocation lines and double diploids) of the thinopyrum intermedium in the distant hybridization process of the wheat and the thinopyrum intermedium. The method has important value for theoretical research and practical application of wheat genetic breeding.

Description

technical field [0001] The invention relates to the technical field of plant molecular genetics and breeding, and more specifically relates to a method for quickly identifying the 4St chromosome of Thipotinum intermedia. Background technique [0002] Since the 1990s, the per unit area yield of wheat in the world has been stagnant. The main reason is that the long-term use of a small number of main cultivars as parents in wheat breeding has resulted in narrower and narrower genetic bases. The disease resistance, stress resistance, and insect resistance of the varieties gradually decline, which reduces the ability of the existing varieties to resist disaster risks. As an important carrier of genes, germplasm resources are called "chips" in the agricultural field. With the increasingly fierce competition for genetic resources in various countries, the shortage of germplasm resources has become the main technical bottleneck restricting the development of the seed industry and th...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13
Inventor 张晓军乔麟轶李欣贾举庆郭慧娟张树伟常利芳陈芳畅志坚
Owner SHANXI AGRI UNIV
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