Molecular marker and method for identifying interspecific hybrids between Chinese cabbages and Brassica carinata and tracking segregation of A02 chromosomes and C02 chromosomes in offspring materials of interspecific hybrids

A molecular marker and chromosome technology, applied in the field of genetics and breeding, can solve the problem of time-consuming and laborious, achieve the effect of simple and fast cost, enrich daily dietary nutrition, and reduce manpower

Active Publication Date: 2019-11-26
INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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  • 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 artifi...

Method used

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  • Molecular marker and method for identifying interspecific hybrids between Chinese cabbages and Brassica carinata and tracking segregation of A02 chromosomes and C02 chromosomes in offspring materials of interspecific hybrids
  • Molecular marker and method for identifying interspecific hybrids between Chinese cabbages and Brassica carinata and tracking segregation of A02 chromosomes and C02 chromosomes in offspring materials of interspecific hybrids
  • Molecular marker and method for identifying interspecific hybrids between Chinese cabbages and Brassica carinata and tracking segregation of A02 chromosomes and C02 chromosomes in offspring materials of interspecific hybrids

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 This example identifies the hybrid F between Chinese cabbage and Ethiopian mustard 1 plant

[0039] 1.1 Extract the F to be detected 1 Genomic DNA of the plant and its parents.

[0040] 1.2 Synthetic primers:

[0041] C02B-F: 5'-GCTTTGTGGGTTTGAGCTTG-3';

[0042] C02B-R: 5'-GCCATTTTGAACCATGAACC-3'.

[0043] 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 + ), 5ng template DNA, 0.2mM dNTPs, 0.5μM primer C02B-F, 0.5μM primer C02B-R, 1U Taq enzyme. PCR reaction conditions: 95°C for 3min; 94°C for 30s, 55.8°C for 30S, 72°C for 30S, 35 cycles; 72°C for 10min.

[0044] 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 electrophoresis until bromophenol blue runs o...

Embodiment 2

[0051] Example 2 This example identifies the hybrid backcross progeny between Chinese cabbage and Ethiopian mustard (BC 2 )Material

[0052] 1.1 Extract the genomic DNA of the plants to be tested and their parents.

[0053] 1.2 Synthetic primers: C02B-F: 5'-GCTTTGTGGGTTTGAGCTTG-3'; C02B-R: 5'-GCCATTTTGAACCATGAACC-3'.

[0054] 1.3PCR 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 15 μL, including: 1×PCR Buffer (containing Mg + ), 1ng template DNA, 0.2mM dNTPs, 0.5μM primer C02B-F, 0.5μM primer C02B-R, 1U Taq enzyme. PCR reaction conditions: 94 3min; 9430s, 55.8°C 30S, 72°C 30S, 35 cycles; 72°C 5min.

[0055] 1.4 The above PCR products were detected by polyacrylamide gel electrophoresis. Configure 8% polypropylene gel, electrophoresis at 180 volts for 2 hours, and end the electrophoresis until bromophenol blue runs out of the bottom of the electr...

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Abstract

The invention discloses a molecular marker and method for identifying interspecific hybrids between Chinese cabbages and Brassica carinata and tracking segregation of A02 chromosomes and C02 chromosomes in offspring materials of the interspecific hybrids, and belongs to the field of plant genetic breeding. The marker comprises a pair of codominant molecular marker bodies, and can be used for identifying the authenticity of the interspecific hybrids between Chinese cabbages and Brassica carinata, and can also be used for identifying and tracking selfing offsprings, backcross offsprings, chromosome addition lines and recombined segregation populations of distant hybrids and segregation of A02 chromosomes and C02 chromosomes in plants.

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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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 张晓辉李锡香宋江萍王海平
Owner INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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