Molecular marker closely linked to peach weeping gene and application of molecular marker

A molecular marker and weeping technology, which is applied in the determination/inspection of microorganisms, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of large genetic distance and difficulty in realizing the molecular identification of peach weeping traits, and achieves simple and convenient methods. Good application promotion prospect, less restrictive effect

Active Publication Date: 2018-07-06
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

Dirlewanger E (1994) discovered the RAPD markers for the weeping trait (pl), and the genetic distances were 11.4 cM (Centimorgan) and 17.2 cM, but the genetic distances were too large to be an accurate marker, and it was difficult to achieve early detection of peach weeping. Molecular Characterization of Clad Traits

Method used

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  • Molecular marker closely linked to peach weeping gene and application of molecular marker
  • Molecular marker closely linked to peach weeping gene and application of molecular marker
  • Molecular marker closely linked to peach weeping gene and application of molecular marker

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Example 1: Identification of weeping branch phenotypes in hybrid segregation populations

[0040] According to the uprightness and degree of weeping branches of peach tree branches, phenotypic evaluation is carried out, wherein the new slightly drooping branches are like weeping willows, which are weeping branches, such as figure 1 Shown; the new branch grows slightly negatively upwards as a common type, such as figure 2 shown.

[0041] "Xinxiang 144" was used as the parent to self-cross, and 152 seedlings of the self-crossed offspring were obtained, and the phenotype evaluation was carried out in April 2016 after planting.

[0042] Through the phenotypic evaluation of the offspring individual plants, it was found that 96 common plants and 56 weeping plants were found. The ratio of the two was close to 3:1, and the P value was 0.700, which conformed to the Mendelian inheritance law, and the weeping traits were controlled by a recessive single gene.

Embodiment 2

[0043] Embodiment two: the extraction of genomic DNA

[0044] Using the CTAB method to extract the genomic DNA of peach leaves, the specific steps are as follows:

[0045] (1) Take about 30 mg of young peach leaves, put them into a 1.2 mL eight-row centrifuge tube, add a 5 mm steel ball to each well, put them into a 96-well base, and freeze them fully with liquid nitrogen;

[0046] (2) Manually shake it several times to ensure that the steel balls fully break the sample, and then grind it with a DNA automatic grinder (Shanghai Lingcheng Biotechnology Co., Ltd.), with a frequency of 30 Hz and a time of 90 s;

[0047] (3) Add 600 μL of the prepared CTAB solution with a 300 mL range 8-channel pipette, put it in a 60°C electric constant temperature blast drying oven and heat for 30 min, during which, shake gently every 10 min;

[0048] (4) Place in a refrigerated centrifuge (Eppendorf 5810 R) at 4°C and centrifuge at 4000 rpm; add a mixture of chloroform and isoamyl alcohol at a ...

Embodiment 3

[0052] Example 3: Primer Design for SNP Development

[0053] Refer to the peach genome sequence (Genome Database for Rosaceae PeachVersion 2.0), and use Primer 3 Input (version4.0) (http: / / primer3.ut.ee / ) to design primers. The length of the primers was 20-25 bp. The SNP markers based on Sanger sequencing were developed, and a pair of primers were designed for every 1 Mb. The length of the amplified fragment was about 700-1600 bp.

[0054] Detailed primer information is shown in Table 1:

[0055] Table 1 Primer information using closely linked SNP markers

[0056] .

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Abstract

The invention discloses a molecular marker closely linked to the peach weeping gene and application thereof the molecular marker, and is expected to be applied to an assistant selection of a peach weeping molecular marker and the breeding of an excellent peach germplasm resource. Through the invention, an SNP molecular marker closely linked to the peach weeping gene is screened and obtained, including WESNP-20.83 and WESNP 20.99, wherein the WESNP-20.83 is located at 20.839 Mb of peach genome Pp03, and the nucleotide of the WESNP-20.83 is G or C; the WESNP-20.99 is located at 20.998 Mb of thepeach genome Pp03, and the nucleotide of the WESNP-20.99 is C or T; and an InDel marker WEInDel-21.61 closely linked to a weeping gene is further obtained. The invention provides a method for assisting in screening of the excellent peach germplasm resource based on developed molecular marker primers; molecular identification and screening of peach weeping traits in an early stage can be realized.The method has the advantages of high efficiency, less restriction and high accuracy.

Description

technical field [0001] The invention relates to the technical field of molecular marker-assisted selection of peach weeping branch, in particular to a molecular marker closely linked with peach weeping branch gene and its application. Background technique [0002] Peach[ Prunus persica (L.) Batsch] is an important deciduous fruit tree native to my country, with a cultivation history of more than 3,000 years; in my country, peach is the third largest deciduous fruit tree, widely cultivated all over the country, and is an important part of agricultural production. [0003] There are many kinds of peach tree growth types, which can be stably inherited, such as dwarf type (DW, dwdw), semi-dwarf type (SD, semi-dwarf), columnar type (PI, brbr), compact type (CT, Ct ) and weeping tree types (WE, plpl), the development and application of new tree types is an important part of my country's breeding work. Among them, dwarf, columnar, and compact tree shapes are of great significance...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 鲁振华王志强张南南牛良崔国朝潘磊曾文芳
Owner ZHENGZHOU FRUIT RES INST CHINESE ACADEMY OF AGRI SCI
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