Molecule marking method for identifying cytoplasm fertility of cayenne pepper in seedling stage

A molecular marker, cytoplasmic technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of long breeding cycle, few sterile sources, limited application, etc., to speed up the breeding process and accelerate the breeding process. Effect

Inactive Publication Date: 2011-08-03
NORTHWEST A & F UNIV
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AI Technical Summary

Problems solved by technology

However, the application of NIC male sterile line in pepper is limited by many unfavorable factors.
First of all, the distribution of fertility restoration genes is not widespread, especially in lantern-type pepper types, and the degree of freedom in combination is very low, which greatly limits its application and is the biggest obstacle to the application of sterile lines at present.
Secondly, there are few sources of sterility and the breeding cycle is long

Method used

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  • Molecule marking method for identifying cytoplasm fertility of cayenne pepper in seedling stage
  • Molecule marking method for identifying cytoplasm fertility of cayenne pepper in seedling stage
  • Molecule marking method for identifying cytoplasm fertility of cayenne pepper in seedling stage

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] 1. Artificially design and synthesize a pair of specific nucleotide sequences as primers, wherein:

[0027] Forward primer: 5'-CCCACACAAGACACATCACA-3'

[0028] Reverse primer: 5'-CGTACGAAATTCTTGGGCTTA-3'

[0029] 2. Extraction of pepper mitochondrial DNA: high salt-proteinase K method (Liu Guangguang et al., Mitochondrial differential gene acquisition and SNP analysis of sterile and maintainer lines of pepper [J], Journal of China Agricultural University, 2010, 15(5): 19~24) Mitochondrial DNA was extracted from 6 pepper lines T130, P65, GS102, PB3, B40, and L33, respectively.

[0030] 3. Perform PCR amplification on the mitochondrial DNA of the above 6 pepper strains with the primers in step 1, and the amplified products are subjected to 0.8% agarose gel electrophoresis, see figure 2 , T130, GS102, L33 plants can appear a specific band with a length of 270bp, which is identified as sterile cytoplasm; while P65, PB3, B40 plants do not have this band, and are identifie...

Embodiment 2

[0036] 1. Artificially design and synthesize a pair of specific nucleotide sequences as primers, wherein:

[0037] Forward primer: 5'-CCCACACAAGACACATCACA-3'

[0038] Reverse primer: 5'-CGTACGAAATTCTTGGGCTTA-3'

[0039] 2. Extraction of capsicum mitochondrial DNA: high salt-proteinase K method (Liu Guangguang et al., Acquisition of mitochondrial differential genes and SNP analysis of sterile line and maintainer line of pepper [J], Journal of China Agricultural University, 2010, 15 (5): 19~24) Mitochondrial DNA of 6 pepper lines P121, P134, K12, K35, FG42, M10 were extracted respectively.

[0040] 3. Using the specific nucleotide sequence described in step 1 as a primer, carry out PCR amplification on the mitochondrial DNA of the above-mentioned 6 pepper strains, and the amplified products are subjected to 0.8% agarose gel electrophoresis, see image 3 , P134, K35, FG42 plants can appear a specific band with a length of 270bp, identified as sterile cytoplasm; while P121, K12, M...

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Abstract

The invention discloses a molecule marking method for identifying the cytoplasm fertility of cayenne peppers in seedling stage, which comprises the following steps of: firstly artificially designing and synthetizing a pair of specific nucleotide sequences as primers; carrying out PCR (Polymerase Chain Reaction) amplification on mitochondria DNA (Deoxyribonucleic Acid) of the cayenne peppers by using the primers; carrying out electrophoresis on amplification products by using 0.8% of agarose gel; if a specific strip appears in the electrophoresis result, representing that the amplification products in the electrophoresis channel are sterile cytoplasm plants; and if no specific strip appears in the electrophoresis result, representing that the amplification products in the electrophoresis channel are fertile cytoplasm plants. The molecule marking method has simple and rapid steps, has stability and reliability, is especially suitable for CMS male sterile molecule marking assistant breeding of the cayenne peppers, and can improve selection efficiency well and quicken the breeding progress, and the identification result is accurate and consistent with the field floral organ fertility identification result and has good repeatability.

Description

technical field [0001] The invention belongs to the field of capsicum breeding and relates to a plant molecular marker method, in particular to a molecular marker-assisted breeding method for identifying the cytoplasmic fertility of peppers at the seedling stage. Background technique [0002] Capsicum is an important cross-pollinated vegetable crop with obvious heterosis. At present, my country's pepper hybrids have occupied a major position in the production, and the main popularized varieties are the first-generation hybrids between varieties or inbred lines. Although this type of hybrid seed has a great potential for increasing production, it must be artificially detasseled during seed production, which is labor-intensive and time-consuming. Moreover, due to the hermaphroditism of peppers, it is difficult to guarantee the purity of the seeds and the cost is relatively high. The utilization of the male sterility trait of pepper solves the problem of emasculation in artifi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 巩振辉黄炜吉姣姣逯明辉陈儒钢
Owner NORTHWEST A & F UNIV
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