Method for detecting chloroplast DNAs to identify cytoplasmic male sterile line of hot pepper

A male sterile line and male sterile technology, applied in the field of detecting chloroplast DNA to identify pepper cytoplasmic male sterile lines, can solve the problems of limiting the development of pepper hybrid breeding, few, difficulty and the like

Inactive Publication Date: 2015-04-15
HUNAN UNIV OF ARTS & SCI +1
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Problems solved by technology

Molecular markers of cytoplasmic male sterile lines have been established in many plants, and the markers can be used to quickly distinguish sterile and fertile materials; studies on the genetic material of pepper male sterile lines are mainly concentrated in the nucleus, and rarely in the cytoplasm

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  • Method for detecting chloroplast DNAs to identify cytoplasmic male sterile line of hot pepper
  • Method for detecting chloroplast DNAs to identify cytoplasmic male sterile line of hot pepper
  • Method for detecting chloroplast DNAs to identify cytoplasmic male sterile line of hot pepper

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Embodiment 1

[0027] Such as Figure 1 to Figure 3 As shown, the present invention takes 3 conventional capsicum varieties as a control, carries out the amplified sequences of chloroplast C1 and C2 fragments on 3 cytoplasmic male sterile lines and corresponding maintainer lines, and uses relevant software analysis to establish capsicum cytoplasmic male sterile Chloroplast DNA molecular fingerprint of the lineage.

[0028] Its specific operation is as follows:

[0029] The first step is to prepare intact chloroplasts, and prepare intact chloroplasts in pepper leaves by differential centrifugation and sucrose density gradient centrifugation;

[0030] The second step is to extract the DNA in the above-mentioned complete chloroplast, and adopt the CTAB method to extract the total DNA in the chloroplast of the pepper leaf to be tested;

[0031] The third step, PCR amplification, carries out PCR amplification to the above-mentioned total DNA with chloroplast primers C1 and C2;

[0032] The fou...

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Abstract

The invention discloses a method for detecting chloroplast DNAs to identify cytoplasmic male sterile line of hot pepper. The method comprises the following steps: taking the leaves of hot pepper as a material, extracting total DNAs in the leaves, carrying out PCR amplification by using chloroplast primers (C1 and C2), carrying out clone sequencing on amplification products, and then making a comparison with chloroplast DNA molecular fingerprints of the cytoplasmic male sterile line of hot pepper, namely identifying the cytoplasmic male sterile line of hot pepper simply, efficiently and accurately. The method for detecting chloroplast DNAs to identify the cytoplasmic male sterile line of hot pepper disclosed by the invention, the clone sequencing is carried out on the PCR amplification products of the chloroplast primers (C1 and C2) only when the cytoplasmic male sterile line of hot pepper is identified, and then a comparison is made, so that the cytoplasmic male sterile line of hot pepper can be identified simply, efficiently and accurately.

Description

technical field [0001] The invention relates to a method for detecting chloroplast DNA to identify cytoplasmic male sterile lines of peppers, belonging to the technical field of cytoplasmic male sterile detection. Background technique [0002] Cytoplasmic male sterility is a common phenomenon in the plant world. The use of male sterility for hybrid breeding has great advantages, such as no need for artificial emasculation, improving the purity of hybrid seeds and increasing yield; the mechanism of cytoplasmic male sterility is very complicated. No two cytoplasmic male sterility mutations were found to be identical. Molecular markers of cytoplasmic male sterile lines have been established in many plants, and the markers can be used to quickly distinguish sterile and fertile materials; studies on the genetic material of pepper male sterile lines are mainly concentrated in the nucleus, and rarely in the cytoplasm According to the report of Pepper Cytoplasmic Male Sterile Lines...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6869C12Q1/6895
Inventor 肖辉海郝小花
Owner HUNAN UNIV OF ARTS & SCI
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