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A method for early detection of peach fruit shape based on chromosome structure variation

A fruit and purpose technology, applied in biochemical equipment and methods, microbiological determination/testing, DNA/RNA fragments, etc.

Active Publication Date: 2022-05-17
BEIJING AGRO BIOTECH RES CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage of this method is that there may be technical problems such as false negatives, such as whether the deletion of the target band is caused by reasons such as poor DNA quality

Method used

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  • A method for early detection of peach fruit shape based on chromosome structure variation
  • A method for early detection of peach fruit shape based on chromosome structure variation
  • A method for early detection of peach fruit shape based on chromosome structure variation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Primer Design for Detecting Peach Fruit Shape

[0035] With the help of the advanced third-generation sequencing platform Pacbio, the present invention completed the genome sequencing of Ruiyoupan 1, and successfully assembled the first draft genome of the flat peach. By comparing the sequence with the existing peach reference genome Lovell (circle), screening the chromosome structure variation, and combining the resequencing data of 149 cultivated peaches, it was found that a chromosomal inversion structure variation located on chromosome 6 was related to the round and flat shape of peach fruit The shape is closely related, and the structural variant was named INV56 (Scaffold_6:27,959,880-29,634,101). Among them, the structural state that exists specifically in flat peach is called INV56F (flat shape, see figure 1 ), another state is INV56R (round, see figure 1 ). Because the locus is heterozygous in flat peaches, the genotype of flat peaches is INV56F / R;...

Embodiment 2

[0039] Embodiment 2: application embodiment

[0040] 137 young leaves of cultivated peaches were collected, including 37 flat peaches and 100 round peaches (Table 2). Genomic DNA was extracted using a plant DNA extraction kit (Tiangen Biochemical Technology (Beijing) Co., Ltd.), and then quantified using sterile Bacteria water was configured as a 20ng / μL template solution. The configuration and reaction conditions of the PCR reaction solution are as follows:

[0041] Composition of PCR reaction solution:

[0042]

[0043]

[0044] PCR reaction conditions:

[0045]

[0046] In the above experimental process, first use the P1 and P2 primer pair for amplification detection. If no specific amplification band is obtained with the P2 primer, continue to use the P3 primer pair for amplification detection. If there are still samples without amplified bands , continue to use the P4 primer pair for amplification detection.

[0047] The detection of the amplification product...

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Abstract

The invention relates to a method for early identification of cultivated peach fruit shape based on PCR technology. The PCR detection of the present invention uses 4 pairs of primers, including 1 pair of primers (P1) for identifying INV56F (Flat), and 3 pairs of primers (P2 / 3 / 4) for identifying INV56R (Round). The identification method is to use the peach genomic DNA to be tested as a template, first use P1 and P2 to amplify respectively, and detect the amplified products. If two specific amplification bands of 1391bp and 705bp are obtained, the fruit shape is flat; if only If there is a 705bp specific amplification band, the fruit shape is round; if no 705bp band is obtained, continue to amplify with the P3 primer pair, if a 981bp specific amplification product band is obtained, the fruit shape is round, otherwise Continue to amplify with the P4 primer; if a 651bp specific amplification product band is obtained, the fruit shape is round, otherwise the fruit shape cannot be judged temporarily. The method has high primer specificity, and the setting of control primers can effectively prevent the occurrence of false negatives, and has the characteristics of simple operation, rapid identification, and accurate results.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the detection of peach fruit shape, and more particularly relates to an early detection method of peach fruit shape based on chromosome structure variation. [0002] technical background [0003] Peach is widely grown in temperate and subtropical regions and has important economic value. Fruit shape is one of the most notable agronomic features of peaches, which can be divided into round peaches and flat peaches. Flat peach, also known as flat peach, is a variety of peach. It has a high edible proportion, sweet and juicy flavor, and is deeply loved by Chinese consumers. In recent years, its proportion in the peach consumption market is gradually increasing. Pantao originated in my country and has a long history. In the middle of the 18th century, pantao was introduced to the West and quickly won the favor of local consumers. It has a large market space. Compared with rou...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156C12Q2600/13
Inventor 谢华徐摇光官健涛于洋
Owner BEIJING AGRO BIOTECH RES CENT
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