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InDel marker for identifying radish flower color control gene and application thereof

A radish and flower color technology, applied in the field of molecular biology, can solve the problems of long breeding cycle, heavy workload, low breeding efficiency, etc., achieve the effect of reducing amplification time and improving detection efficiency

Active Publication Date: 2021-07-23
INST OF ECONOMIC CROP HUBEI ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Using traditional breeding methods to obtain the corresponding varieties of traits that people care about is heavy workload, low breeding efficiency and long breeding cycle

Method used

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  • InDel marker for identifying radish flower color control gene and application thereof
  • InDel marker for identifying radish flower color control gene and application thereof
  • InDel marker for identifying radish flower color control gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] F 1 , F 1 F2 color segregation population obtained by selfing ( figure 1 ). The results of genetic analysis showed that the flower colors of the F1 offspring of "QZ-16" and "55" were all purple. In 309 F 2 In the population, there were 238 purple-flowered plants and 71 white-flowered plants, such as figure 1 As shown, the separation ratio is 3:1(χ 2 =0.67, P=0.42>0.05), in line with the Mendelian inheritance ratio. Therefore, the purple flower trait of radish is dominant inheritance controlled by a single gene.

[0035] In order to identify the Pf gene controlling radish flower color, the target gene controlling radish flower color was identified by map-based cloning method.

[0036] Using BSA combined with genome-wide resequencing (i.e. QTL-seq) for initial mapping of the Pf gene, in F 2 From the segregation population, 20 purple-flowered plants were selected to build the "purple flower pool", and 20 white-flowered plants were selected to build the "white flowe...

Embodiment 2

[0046] Application of InDel mark

[0047] Utilize the InDel marker identification among the embodiment 1 to confirm the color (white / purple) of radish flower, the steps are as follows:

[0048] Select radish varieties and materials

[0049] Four radish cultivars bred by the Hubei Provincial Key Laboratory of Vegetable Germplasm Innovation and Genetic Improvement and three commercially available radish high-generation inbred lines were used to verify the flower color phenotype. The specific steps are as follows:

[0050] Take about 2cm of plant leaves and put them into a 2mL centrifuge tube;

[0051] Add 1 or 2 abrasive steel balls and 800 μL of 1.5% CTAB extract;

[0052] Use a sample grinder to grind the sample for 90s, and the sample grinding frequency is 60Hz;

[0053] Water bath at 65°C for 30 minutes, and shake upside down 3 times during the period;

[0054] Add 600 μL of 24:1 (chloroform:isoamyl alcohol, volume ratio), mix up and down for 5 minutes, until the lower s...

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Abstract

The invention provides an InDel molecular marker for identifying a radish flower color control gene and application thereof. A forward primer sequence of the InDel molecular marker is as shown in SEQ ID NO.4, reverse primers comprises a first reverse primer and a second reverse primer, a sequence of the first reverse primer is as shown in SEQ ID NO.5, and a sequence of the second reverse primer is as shown in SEQ ID NO.6. Compared with the prior art, the InDel molecular marker has the beneficial effects that the radish flower color can be detected in the seedling stage by detecting the deletion or insertion of the provided specific fragment, and the InDel molecular marker can be used for subsequent identification and breeding.

Description

technical field [0001] The invention belongs to the field of molecular biology, and in particular relates to an InDel molecular marker for identifying radish flower color control genes and its application. Background technique [0002] Radish (Raphanus sativus L.) is one of the important staple vegetables in my country, belonging to the genus Radish in the Brassicaceae vegetable crop. Radish flowers vary in variety, and can be divided into white, purple, red, yellow-green and so on. The driving force of radish petal color variation mainly comes from the attraction of pollinators, which in turn affects its own pollination efficiency and seed setting rate, which is one of the important agronomic traits of radish. Using traditional breeding methods to obtain the corresponding varieties of traits that people care about requires a lot of work, low breeding efficiency and long breeding cycle. [0003] With the development of molecular biology, molecular marker-assisted selection...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/124C12Q2600/156C12Q2600/172
Inventor 严承欢邱正明黄燕崔磊邓晓辉矫振彪刘志雄张振兴朱凤娟
Owner INST OF ECONOMIC CROP HUBEI ACADEMY OF AGRI SCI
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