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Primer combination for identifying authenticity of hybrid seeds of rape and radish and application of primer combination

A primer combination and hybrid technology, applied in the field of agricultural molecular biology, can solve rare problems and achieve the effects of good repeatability, good versatility, accurate and rapid identification

Pending Publication Date: 2022-05-06
JIANGSU COASTAL AREA AGRI SCI RES INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

SSR markers have become a common method for identifying seed purity at this stage, but there are few reports on using SSR molecular marker technology to identify the authenticity of rapeseed and radish hybrids

Method used

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  • Primer combination for identifying authenticity of hybrid seeds of rape and radish and application of primer combination
  • Primer combination for identifying authenticity of hybrid seeds of rape and radish and application of primer combination
  • Primer combination for identifying authenticity of hybrid seeds of rape and radish and application of primer combination

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] 1. Brassica napus, radish and its hybrid F 1 Germination and culture of generation seeds

[0048] Select healthy and plump Brassica napus and radish and their hybrid F 1 Substitute seeds were washed with distilled water, placed in a moistened glass petri dish with two layers of filter paper, and placed in a light incubator at 28°C to germinate, with or without light. After 3-4 days, the germinated seeds are transferred to the water-absorbing sponge holes to continue culturing, and can be harvested after 15 days for the extraction of genomic DNA.

[0049] The seed germination rates of the three samples were all above 80%, and the parents and their hybrids were all successfully cultivated into seedlings, wherein harvesting each 4g of the parent plants and 8 strains of F 1 generation hybrids.

[0050] Female parent, male parent and their hybrid F 1 The germination period of generation seeds is shown in the figure Figure 1-3 shown.

[0051] 2. Brassica napus, radish ...

Embodiment 2

[0076] Embodiment 2 primers and parent, hybridization F 1 Generation PCR amplification

[0077] Utilize the primer that embodiment 1 screens out to carry out hybrid progeny identification, concrete steps are as follows:

[0078] (1) Extraction of genomic DNA of rapeseed, radish and varieties to be tested:

[0079] Genomic DNA was extracted using the Biospin Plant Genomic DNA Extraction Kit from Hangzhou Bioer, the product number is BSC13S1, and the specific method can be found in the manual of the extraction kit.

[0080] (2) Using the genomic DNA extracted in step (1) as a template respectively, carry out PCR amplification with Primer17, Primer51, Primer125, Primer128 and Primer140 primer combinations to obtain the amplified product; the specific parameters of the PCR reaction are the same as in Example 1;

[0081] (3) The amplified product is subjected to polyacrylamide gel electrophoresis to obtain an electrophoretic spectrum; the specific processing parameters are the sa...

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Abstract

The invention discloses a primer combination for identifying the authenticity of rape and radish hybrids and application of the primer combination. Belongs to the technical field of agricultural molecular biology. Primer17, Primer51, Primer125, Primer128 and Primer140 are screened out from a large number of SSR primers to form a primer combination for identifying the authenticity of the rape and radish hybrids, PCR amplification is conducted through genomic DNA of parents and the hybrids, an electrophoretogram is obtained through gel electrophoresis treatment, and the authenticity of the rape and radish hybrids is identified. The authenticity of the to-be-detected hybrids can be judged by observing whether the hybrids have the characteristic bands of the parents or not, and the method is convenient, rapid and simple in operation step, can be used for rapidly identifying a large number of to-be-detected hybrids, and is accurate in determination result.

Description

technical field [0001] The invention relates to the technical field of agricultural molecular biology, in particular to a combination of primers for identifying the authenticity of rapeseed and radish hybrids and an application thereof. Background technique [0002] Distant hybridization is an important way to create new varieties of crops, and it is also a powerful tool for improving species. The use of distant hybridization technology can break the isolation between species and genus, broaden the genetic basis and create new germplasm for breeding programs, so as to effectively transfer the target genetic material between species and genus. Distant hybridization has been widely used in rapeseed breeding and achieved remarkable results. At present, although there are reports on the distant hybridization and hybrid identification of Brassica napus and radish at home and abroad, there are only a few. The hybridization between Brassica napus and radish belongs to the distant...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6895C12Q1/6858C12Q2600/156C12Q2531/113C12Q2565/125
Inventor 万林生孙红芹倪正斌韩配配陈志爱孙明法严国红朱国永王爱民唐红生刘凯施伟孙一标张桂云代金英朱静雯赵绍路胡蕾程新杰张梦龙岳红亮
Owner JIANGSU COASTAL AREA AGRI SCI RES INST
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