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Molecular markers for identification of interspecific hybrids of Chinese kale and red brassica campestris and tracing of chromosome segregation conditions of progeny materials A05 and C04 of interspecific hybrids

A technology of molecular markers and chromosomes, applied in the field of genetic breeding, can solve the problems of time-consuming and laborious, and achieve the effect of simple and fast cost, simple operation, and rich vegetable types

Active Publication Date: 2019-11-08
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 for identification of interspecific hybrids of Chinese kale and red brassica campestris and tracing of chromosome segregation conditions of progeny materials A05 and C04 of interspecific hybrids
  • Molecular markers for identification of interspecific hybrids of Chinese kale and red brassica campestris and tracing of chromosome segregation conditions of progeny materials A05 and C04 of interspecific hybrids
  • Molecular markers for identification of interspecific hybrids of Chinese kale and red brassica campestris and tracing of chromosome segregation conditions of progeny materials A05 and C04 of interspecific hybrids

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 This example identifies the hybrid F between Chinese kale and red cabbage 1 Plant

[0043] 1.1 Extract F to be detected 1 Genomic DNA of the plant and its parents.

[0044] 1.2 Synthetic primers:

[0045] C04a-F: 5'-TCCGGAACTCATGAATGCTT-3' (SEQ ID No. 1);

[0046] C04a-R: 5'-TTTTGGGTGCCTCGTGTAAT-3' (SEQ ID No. 2).

[0047] 1.3 PCR amplification. To be tested F 1 The plant and its parental DNA are used as templates, and the above primers are used for PCR amplification reactions. The reaction system is 10μL, including: 1×PCR Buffer (containing Mg + ), 0.5ng template DNA, 0.2mM dNTPs, 0.5μM primer C04a-F, 0.5μM primer C04a-R, 1U Taq enzyme. PCR reaction conditions: 94℃3min; 94℃30s, 55.8℃30S, 72℃30S, 35 cycles; 72℃10min.

[0048] 1.4 The above PCR products were detected by polyacrylamide gel electrophoresis. Configure 8% polypropylene gel, electrophoresis at 180 volts for 1.5 hours, until bromophenol blue runs out of the bottom of the electrophoresis tank to end electrop...

Embodiment 2

[0055] Example 2 In this example, the backcross progeny of interspecific hybrids between Chinese kale and red cabbage (BC 1 )material

[0056] 1.1 Extract the genomic DNA of the plant to be tested and its parents.

[0057] Synthesis primer:

[0058] C04a-F: 5'-TCCGGAACTCATGAATGCTT-3' (SEQ ID No. 1);

[0059] C04a-R: 5'-TTTTGGGTGCCTCGTGTAAT-3' (SEQ ID No. 2).

[0060] 1.3 PCR amplification. Using the DNA of the plant to be tested and its parental DNA as a template, the PCR amplification reaction is carried out with the above primers. The reaction system is 20μL, including: 1×PCR Buffer (containing Mg + ), 1ng template DNA, 0.2mM dNTPs, 0.5μM primer C04a-F, 0.5μM primer C04a-R, 1U Taq enzyme. PCR reaction conditions: 95℃3min; 95℃30s, 55.8℃30S, 72℃30S, 35 cycles; 72℃5min.

[0061] 1.4 The above PCR products were detected by polyacrylamide gel electrophoresis. Configure 8% polypropylene gel, electrophoresis at 180 volts for 2 hours, until bromophenol blue runs out of the bottom of the el...

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Abstract

The invention provides molecular markers and an identification method for identification of interspecific hybrids of Chinese kale and red brassica campestris and chromosome segregation conditions of progeny materials A05 and C04 of the interspecific hybrids, and belongs to the field of plant genetics and breeding. According to the molecular markers and the identification method for identificationof the interspecific hybrids of the Chinese kale and the red brassica campestris and the chromosome segregation conditions of the progeny materials A05 and C04 of the interspecific hybrids, the markers are a pair of codominant molecular markers, and the markers are used for identifying the interspecific crossing true hybrids between the Chinese kale and the red brassica campestris, and can also beused for identifying selfing progeny, backcross progeny, a chromosome addition line and a recombination segregation population and plant of a distant hybrid.

Description

Technical field [0001] The present invention relates to the field of genetic breeding, in particular to methods 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 species reproductive isolation, distant hybridization often requires techniques such as artificial pollination and embryo rescue, which are time-consuming and laborious, and require certain scientific training. In the process of distant hybridization, false hybrid plants may be produced due to incomplete emasculation, the development of female gametes into plants, and other reasons. Therefore, plants obtained by distant hybridization need to be tested by molecular and cytological methods to determine whether they are true hybrids. The selfing and backcrossing offspring of distant hybrids need to track chromosomes or chromosome fragments through molecular or cytological m...

Claims

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Application Information

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Patent Type & Authority Applications(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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