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Group of InDel markers for identifying green peach aphid resistance/susceptibility traits of mountain peach hybrid population and application thereof

An indel-2, mountain peach technology, applied in recombinant DNA technology, microbial determination/inspection, DNA/RNA fragments, etc., can solve the problems of long distance and low accuracy of peach aphid resistance genes

Pending Publication Date: 2021-07-30
ZHENGZHOU FRUIT RES INST CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, the existing linkage markers are far away from the gene of peach aphid resistance, and the accuracy rate is low in the early identification of hybrid offspring

Method used

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  • Group of InDel markers for identifying green peach aphid resistance/susceptibility traits of mountain peach hybrid population and application thereof
  • Group of InDel markers for identifying green peach aphid resistance/susceptibility traits of mountain peach hybrid population and application thereof
  • Group of InDel markers for identifying green peach aphid resistance/susceptibility traits of mountain peach hybrid population and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: Obtaining of Nucleotide Indel (InDel) Markers

[0028] In the present invention, 179 individual plants in the F2 population of rubber nectarine × broom-shaped mountain peach hybrids are used as materials for BSA positioning, and 35 Indels are used to further encrypt, locate to the interval of 950kb, and combine aphid-resistant individual plants and aphid-sensitive individual plants for inoculation In the gene expression data after aphids, Prupe.3G275300 and Prupe.3G262100 were screened, and InDel-1 (between 25387879 and 25387902bp of the third chromosome of the peach genome) and InDel were found on the promoters of these two genes, respectively. -2 (between 24761660 and 24761668 bp of the third chromosome of the peach genome).

Embodiment 2

[0029] Embodiment 2: Utilize InDel mark to identify the method that mountain peach hybrid population is resistant to peach aphid

[0030] 1. DNA extraction

[0031] The modified CTAB method was used to extract the DNA of the mountain peach sample tissue (leaf) to be tested, and the DNA sample volume was not less than 30 μl. Measure the OD value of the DNA sample at 260nm and 280nm with a UV spectrophotometer, and calculate the DNA content and OD 260 / 280 ratio. DNA sample purity OD 260 / 280 The value should be between 1.8-2.0, and the concentration should be diluted to around 10ng / μl.

[0032] 2. Primer design

[0033] InDel-1 and InDel-2 primers were designed according to the upstream and downstream reference sequences of the deletion site (InDel-1 about 170bp, InDel-2 about 690bp), see Table 1 below for details.

[0034] Table 1 Sequence information at both ends of the insertion site

[0035]

[0036] Note: The nucleotide sequence of the gray bottom part is the sequen...

Embodiment 3

[0051] Example 3: Blind test verification of performance of 2 hybrid populations using a set of InDel markers invented

[0052] 1. Selection of test materials

[0053] Using the conventional peach varieties planted in the resource garden of Zhengzhou Pomology Research Institute as experimental materials, 2 mountain peach hybrid populations whose phenotypic traits have been investigated were selected, with a total of 224 individual plants. See Table 5 for details.

[0054] Table 5 The name and population size information of the tested hybrid population

[0055]

[0056] Note: Broom-shaped mountain peach is a wild germplasm resistant to aphids, West 29-13 in South District 1 is an excellent line resistant to aphids, rubber nectarine is a variety bred with aphid susceptibility, and Sweet dream is a variety bred with aphid susceptibility.

[0057] 2. Utilize the identification method of a group of InDel marks of the present invention

[0058] Using a set of InDel markers of t...

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Abstract

The invention discloses a group of InDel markers for identifying green peach aphid resistance / susceptibility traits of mountain peach hybrid population and application thereof, the InDel markers comprise an InDel-1 marker and an InDel-2 marker, the nucleotide sequence of the InDel-1 marker is shown as SEQ ID NO.1, and the nucleotide sequence of the InDel-2 marker is shown as SEQ ID NO.2. Two groups of InDel primer pairs are designed aiming at InDel-1 and InDel-2 markers, electrophoresis detection is performed on a PCR amplification product amplified by the InDel-1 corresponding primer pair, sequencing is performed on a PCR amplification product amplified by the InDel-2 corresponding primer pair, and the peach aphid resistance / infection character of the mountain peach is predicted according to a result. Molecular marker genotype accuracy verification is carried out on 224 single plants to be detected in two hybrid populations, and the result shows that the verification accuracy reaches 96.88%. The nucleotide insertion and deletion (InDel) marker has the advantages of rapidness, convenience, low cost and the like when being used for identifying the resistance of the green peach aphid, and can be applied to production in a large scale.

Description

technical field [0001] The invention relates to a group of InDel markers, a pair of primers and an application thereof for identifying aphid resistance / susceptibility traits of mountain peach hybrid populations, and belongs to the field of biotechnology. Background technique [0002] Peach aphid is an important pest in the production of peach trees. From mid-February to mid-March in spring, the larvae of peach aphid hatched from overwintering eggs can only survive and grow up by eating the leaves of peaches and mountain peaches. Young leaves cause immediate damage. The young leaves of the plants eaten by the green peach aphid are rolled horizontally or irregularly to the opposite side, which hinders the nutrient absorption of the leaves, deteriorates the nutrition of the leaves, and even falls yellow. The honeydew excreted by aphids on the leaves can easily induce mildew and interfere with the photosynthesis of plants; after the young fruit is bitten by the green peach aphi...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/6858C12N15/11
CPCC12Q1/6895C12Q1/6858C12Q2600/13C12Q2600/156C12Q2531/113C12Q2565/125C12Q2535/122
Inventor 王力荣王君秀李勇王新卫朱更瑞曹珂吴金龙陈昌文方伟超
Owner ZHENGZHOU FRUIT RES INST CHINESE ACADEMY OF AGRI SCI
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