Molecular marker method for identifying pepper anti-epidemic disease character

A technology for molecular markers and anti-epidemic diseases, applied in biochemical equipment and methods, microbial measurement/inspection, etc., to achieve simple and fast steps, improved selection efficiency, and good repeatability

Inactive Publication Date: 2009-07-22
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although the primers selected by this method are complementary to known DNA sequences, the 3' ends of the two primers are reversed.

Method used

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  • Molecular marker method for identifying pepper anti-epidemic disease character
  • Molecular marker method for identifying pepper anti-epidemic disease character
  • Molecular marker method for identifying pepper anti-epidemic disease character

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] 1. Artificially designed and synthesized nucleotide sequences as primers:

[0040] Forward primer: 5’-AGGTGGTCTTCAATGATCAGAGA-3’

[0041] Reverse primer: 5’-CCTGAGGCAAATTCCAAATCTC-3’

[0042] 2. Extraction of genomic DNA of pepper: A modified SDS method was used to extract genomic DNA of 7 pepper inbred lines A5, A11, B17, B23, B2, B32 and B19 (Gong Zhenhui, etc., using PCR to identify the trace DNA extraction method of transgenic plants [ J]. Journal of Northwest Agricultural University, 1997, 25(1): 45-47).

[0043] 3. Using the nucleotide molecules described in step 1 as primers, PCR amplification was performed on the genomic DNA of the 7 pepper inbred lines mentioned above. All 7 pepper inbred lines could amplify a fragment of 590bp in length ( figure 1 Agarose gel electrophoresis analysis of PCR amplification products of 7 pepper inbred lines. M: DNA Marker; 1: A5; 2: A11; 3: B17; 4: B23; 5: B2; 6: B32; 7: B19). The PCR amplification system is as follows:

[0044] (1)...

Embodiment 2

[0051] 1. Artificially designed and synthesized nucleotide sequences as primers:

[0052] Forward primer: 5’-AGGTGGTCTTCAATGATCAGAGA-3’

[0053] Reverse primer: 5’-CCTGAGGCAAATTCCAAATCTC-3’

[0054] 2. Extraction of pepper genomic DNA: The genomic DNA of 7 pepper inbred lines B3, P11, B4, P30, B12, P97 and P45 were extracted using modified SDS method.

[0055] 3. Using the nucleotide molecules described in step 1 as primers, PCR amplification was performed on the genomic DNA of the 7 pepper inbred lines mentioned above. All 7 pepper inbred lines could amplify a fragment of 590bp in length ( image 3 Agarose gel electrophoresis analysis of PCR amplification products of 7 pepper inbred lines. M: DNA Marker; 1: B3; 2: P11; 3: B4; 4: P30; 5: B12; 6: P97; 7: P45). The PCR amplification system is as follows:

[0056] (1) The volume of the reaction solution is 25μL, and the composition and content of the articles used are:

[0057] 10 times PCR buffer 2.5μL, 25mmol·L -1 MgCl 2 2.0μL, 5U·...

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Abstract

The invention discloses a molecular marking method that is used for differentiating pepper disease resisting characteristics, which comprises extraction of pepper genomic DNA, PCR argumentation and polymorphism analysis of a specific enzyme incision product, and is characterized in that a pair of nucleotide molecules with a specified sequence that consists of a certain number of basic groups are artificially synthesized according to a plant disease resistance gene sequence and used as a primer, PCR reaction is realized to obtain a molecular marker, and pepper disease resistant materials and pepper disease susceptible materials can be identified through the size of the specific enzyme incision product in the marker. The molecular marking method can be used for disease resistance filtering in pepper seedling or adult phases and can greatly accelerate the process of disease-resistant pepper breeding.

Description

Technical field [0001] The present invention belongs to the field of pepper breeding, and specifically relates to a plant molecular marker technology, in particular to a molecular marker method for identifying disease resistance traits of pepper. The method is suitable for molecular marker assisted breeding for disease resistance. Background technique [0002] Pepper blight is one of the main diseases that endanger pepper production. It occurs in many countries in the world. It is found in major pepper producing areas such as Qinghai, Xinjiang, Gansu, Shaanxi, Beijing, Liaoning, Shanghai, Zhejiang, Hunan, Sichuan, and Yunnan. There are reports of widespread occurrence and epidemics. With the development of the pepper industry, pepper blight tends to increase year by year, causing serious economic losses. The pathogen of the disease is (Phytophthora capsici Leon), which can be affected at the seedling and adult stage of pepper, and the stems, leaves and fruits can be affected. The...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 巩振辉李大伟黄炜贾庆利逯明辉陈儒钢
Owner NORTHWEST A & F UNIV
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