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Molecular markers and methods for identifying chromosome segregation of a06 and c07 in Brassica vegetable interspecific hybrids and progeny materials

A technology of molecular markers and chromosomes, applied in the field of genetic breeding, can solve time-consuming and labor-intensive problems, and achieve the effect of simple and fast cost, low technical requirements, and expansion of genetic resources

Active Publication Date: 2022-04-22
INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the reproductive isolation of species, distant hybridization often requires techniques such as artificial pollination and embryo rescue, which is time-consuming and laborious, and requires certain scientific training

Method used

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  • Molecular markers and methods for identifying chromosome segregation of a06 and c07 in Brassica vegetable interspecific hybrids and progeny materials
  • Molecular markers and methods for identifying chromosome segregation of a06 and c07 in Brassica vegetable interspecific hybrids and progeny materials
  • Molecular markers and methods for identifying chromosome segregation of a06 and c07 in Brassica vegetable interspecific hybrids and progeny materials

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 This example identifies the hybrid F between Chinese cabbage and Ethiopian mustard 1 plant

[0047] 1.1 Extract the F to be detected 1 Genomic DNA of the plant and its parents.

[0048] 1.2 Synthetic primers:

[0049] C07D-F: 5'-GGAGAAGAAAACAGCGATGC-3' (SEQ ID No.1);

[0050] C07D-R: 5'-GGAATAGCTCTTGACGCTCG-3' (SEQ ID No. 2).

[0051] 1.3PCR amplification. To be detected F 1 Plants and their parental DNA are used as templates, and the above primers are used for PCR amplification reaction. The reaction system is 10 μL, including: 1×PCR Buffer (containing Mg 2+ ), 0.5ng template DNA, 0.2mM dNTPs, 0.5μM primer C07D-F, 0.5μM primer C07D-R, 1U Taq enzyme. PCR reaction conditions: 95°C for 3min; 95°C for 30s, 59.8°C for 30S, 72°C for 30S, 35 cycles; 72°C for 10min.

[0052] 1.4 The above PCR products were detected by polyacrylamide gel electrophoresis. Configure 8% polypropylene gel, run electrophoresis at 180 volts for 1.5 hours, and end the electrophores...

Embodiment 2

[0058] Example 2 This example identifies the interspecific hybrid F of Chinese kale and red cabbage moss 1 plant

[0059] 1.1 Extract the F to be detected 1 Genomic DNA of the plant and its parents.

[0060] 1.2 Synthetic primers:

[0061] C07D-F: 5'-GGAGAAGAAAACAGCGATGC-3' (SEQ ID No.1);

[0062] C07D-R: 5'-GGAATAGCTCTTGACGCTCG-3' (SEQ ID No. 2).

[0063] 1.3 PCR amplification. To be detected F 1 Plants and their parental DNA are used as templates, and the above primers are used for PCR amplification reaction. The reaction system is 15 μL, including: 1×PCR Buffer (containing Mg 2+ ), 1ng template DNA, 0.2mM dNTPs, 0.5μM primer C07D-F, 0.5μM primer C07D-R, 1U Taq enzyme. PCR reaction conditions: 94°C for 3min; 94°C for 30s, 59.8°C for 30S, 72°C for 30S, 35 cycles; 72°C for 5min.

[0064] 1.4 The above PCR products were detected by polyacrylamide gel electrophoresis. Configure 8% polypropylene gel, run electrophoresis at 180 volts for 1.5 hours, and end the electropho...

Embodiment 3

[0069] Example 3 This example identifies the hybrid backcross progeny between Chinese cabbage and Ethiopian mustard (BC 2 )Material

[0070] 1.1 Extract the genomic DNA of the plants to be tested and their parents.

[0071] 1.2 Synthetic primers:

[0072] C07D-F: 5'-GGAGAAGAAAACAGCGATGC-3' (SEQ ID No.1);

[0073] C07D-R: 5'-GGAATAGCTCTTGACGCTCG-3' (SEQ ID No. 2).

[0074] 1.3 PCR amplification. Taking the plant to be detected and its parental DNA as a template, the PCR amplification reaction is carried out with the above primers. The reaction system is 20 μL, including: 1×PCR Buffer (containing Mg +), 10ng template DNA, 0.2mM dNTPs, 0.5μM primer C07D-F, 0.5μM primer C07D-R, 1U Taq enzyme. PCR reaction conditions: 94°C for 3min; 94°C for 30s, 59.8°C for 30S, 72°C for 30S, 35 cycles; 72°C for 5min.

[0075] 1.4 The above PCR products were detected by polyacrylamide gel electrophoresis. Configure 8% polypropylene gel, run electrophoresis at 180 volts for 1 hour, and stop ...

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Abstract

The invention provides an SSR molecular marker and a method for identifying the chromosome segregation of A06 and C07 interspecific hybrids of Brassica vegetables and offspring materials, and belongs to the field of plant genetics and breeding. The molecular marker can quickly and accurately identify the interspecific hybrids of Brassica A genome (such as Chinese cabbage, red cabbage, etc.) . The method can identify distant hybrids of Brassica A genome and C genome plants and their backcross offspring, which can expand the genetic resources of Brassica vegetables, transfer new traits, enrich vegetable types, and enrich people's daily dietary nutrition.

Description

technical field [0001] The invention relates to the field of genetic breeding, in particular to a method for identification and selection of distant hybrid plants. Background technique [0002] Distant hybridization is an important means to create new plant germplasm and expand breeding resources. Due to the reproductive isolation of species, distant hybridization often requires techniques such as artificial pollination and embryo rescue, which is time-consuming and laborious, and requires certain scientific training. In the process of distant hybridization, false hybrid plants may be produced due to incomplete castration, female gametes developing into plants, and other reasons. Therefore, the plants obtained by distant hybridization need to be tested whether they are true hybrids by molecular, cytological and other methods. The offspring of self-crossing and backcrossing of distant hybrids need to track chromosomes or chromosome fragments by molecular or cytological meth...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 李锡香张晓辉宋江萍王海平
Owner INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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