Indel molecular marker primer closely linked with wax gourd fruit size and application of Indel molecular marker primer

A technology of molecular markers and wax gourd, which is applied in the field of molecular biology, can solve the problems of high cost and long time consumption, and achieve the effect of low cost, simple operation and strong specificity

Active Publication Date: 2022-04-29
INST OF VEGETABLES GUANGDONG PROV ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Conventional breeding needs to identify the size of the fruit after the fruit is mature, which is time-consuming and costly

Method used

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  • Indel molecular marker primer closely linked with wax gourd fruit size and application of Indel molecular marker primer
  • Indel molecular marker primer closely linked with wax gourd fruit size and application of Indel molecular marker primer
  • Indel molecular marker primer closely linked with wax gourd fruit size and application of Indel molecular marker primer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] In this example, large-fruited wax gourd B227 (obtained from Sanshui black-skinned wax gourd through conventional multi-generation selection) and small-fruited wax gourd HF3 (daibao wax gourd obtained through conventional multi-generation selection) were used as parents to construct a 4-generation segregation population to study fruit size The results showed that in this population, fruit size is a quality trait controlled by a pair of nuclear genes, and large fruit is dominant over small fruit. Subsequently, by F 2 Gene pools were constructed for individual plants of extreme-sized fruits in the population, and BSA-seq was performed to preliminarily locate the fruit-sized genes on chromosome 10. Further construction of F containing 1486 individual plants 2 The population was marked and encrypted, and the fruit size gene was mapped to the interval of about 96.36kb (16809996-16906354) on chromosome 10.

[0034] In fact, in addition to the large-fruited wax gourd B227 an...

Embodiment 2

[0041] The polymorphism obtained in Example 1 is marked at 178 F 2 Amplification verification in individual plants of the population ( figure 2 , image 3 ), including the following steps:

[0042] (1) Extraction of wax gourd DNA

[0043] The experimental materials were large-fruited wax gourd B227 and small-fruited wax gourd HF3 and F 2 For the fresh leaves of a single plant in the population, the steps of extracting genomic DNA are as follows:

[0044] ①Put a small amount of fresh leaves into a 2mL centrifuge tube, add steel balls, grind on a sample mill, shake 30 times per second for 2 minutes, add 800 μL of 2% CTAB extract, mix well, and place in a 65°C water bath for 1 hour (every Shake once every 10 minutes);

[0045] ②After standing at room temperature, add 800 μL of chloroform:isoamyl alcohol (volume ratio 24:1), mix gently for 10 minutes and centrifuge at 12000 rpm for 15 minutes, transfer the supernatant (about 600 μL) to a new 1.5 mL centrifuge tube;

[0046]...

Embodiment 3

[0068] Using the indel molecular marker primer in Example 1 and the method in Example 2, 130 parts of breeding material single plants are verified, and the single plant DNA is extracted for detection, and the results are as follows Figure 4 shown, from Figure 4 It can be seen from the figure that among materials 1-114, only 113 is a large fruit (with FS 168 and FS 173 ), the rest of the materials are small fruits (only with FS 173 ), in materials 115-130, all are large fruit (only with FS 168 ). At the same time, the fruit size phenotype was identified, and the results showed that the marker detection results were consistent with the field phenotype.

[0069] The above examples show that the method of the present invention can effectively distinguish large fruit and small fruit materials, and accurately and quickly detect the fruit size phenotype of the materials.

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Abstract

The invention discloses an Indel molecular marker primer closely linked with the size of a wax gourd fruit, the Indel molecular marker primer comprises an upstream primer BhFS-F and a downstream primer BhFS-R, the nucleotide sequence of the upstream primer BhFS-F is as shown in SEQ ID NO.1, and the nucleotide sequence of the downstream primer BhFS-R is as shown in SEQ ID NO.2. The Indel molecular marker primer has the advantages that the Indel molecular marker primer is closely linked with the size of the wax gourd fruit; the primer can generate a large fruit specific marker BhFS168 of 168bp and a small fruit specific marker BhFS173 of 173bp, and is good in repeatability and strong in specificity. The invention also discloses an application of the Indel molecular marker primer in the aspect of identifying the size phenotype of the wax gourd fruit in the seedling stage. According to the application, the Indel molecular marker primer is utilized, and the size of the wax gourd fruit can be accurately, rapidly and effectively identified.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to an Indel molecular marker primer closely linked with the fruit size of wax gourd and its application. Background technique [0002] Fruit size is an important agronomic trait of wax gourd, which directly determines the yield of wax gourd. Fruit size is an agronomic trait after the fruit is fully mature, and it is easily affected by the environment, which is a difficult point in trait improvement. Conventional breeding needs to identify the size of the fruit after the fruit is ripe, which is time-consuming and costly. By constructing genetic populations, BSA-seq initial positioning, and further using large populations to encrypt markers, fine-mapping and obtaining molecular markers co-segregated with fruit size, the selection efficiency of fruit size can be significantly improved by marker-assisted selection technology. The size of the fruit can be selected...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6895C12Q1/6858C12Q2600/13C12Q2600/156C12Q2531/113C12Q2565/125
Inventor 刘文睿孙飘云闫晋强王毅谢大森江彪王敏杨松光罗陈史绍琪
Owner INST OF VEGETABLES GUANGDONG PROV ACAD OF AGRI SCI
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