Molecular marker primer composition for identifying freestone trait of prunus persica fruits and application of molecular marker primer composition

A primer combination and molecular labeling technology, applied in the field of molecular biology, can solve the problems that the last single base cannot achieve the resolution effect and the primer cannot be effectively extended, and achieve the effect of reducing the test cost, reliable detection effect and cost reduction.

Pending Publication Date: 2019-12-06
NINGBO ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Theoretically, as long as the last base is not paired, the primer cannot be effectively extended; but the last single base mismatch often cannot achieve the expected resolution effect

Method used

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  • Molecular marker primer composition for identifying freestone trait of prunus persica fruits and application of molecular marker primer composition
  • Molecular marker primer composition for identifying freestone trait of prunus persica fruits and application of molecular marker primer composition
  • Molecular marker primer composition for identifying freestone trait of prunus persica fruits and application of molecular marker primer composition

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] A set of application methods for identifying the molecular marker primer combination of peach sticky core traits, the detailed steps are:

[0048] (1) Primer synthesis: According to the sequence information in Table 1, Sangon Bioengineering (Shanghai) Co., Ltd. was entrusted to synthesize primers JYSNP-1, JYSNP-2, JYSNP-3 and tail primers respectively; wherein, the 5' end of the tail primers Add three different fluorophores, namely FAM, HEX and NED;

[0049] Table 1: Related Primer Sequences

[0050]

[0051] Note: * indicates thio modification.

[0052] (2) Extraction of peach leaf DNA of the population sample to be tested:

[0053] A. Grind about 0.2g of young leaves with liquid nitrogen into powder and place them in a 2mL centrifuge tube;

[0054] B. Add 0.8mL CTAB lysate preheated at 65°C and mix well. The formula of the lysate is: 100mmol / L Tris Cl (pH 8.0), 20mmol / L EDTA (pH 8.0), 1.4mmol / L NaCl, 2.5% CTAB (M / V), 3% PVP (M / V), 2% β-mercapto...

Embodiment 2

[0067] A set of application methods for identifying the molecular marker primer combination of peach sticky core traits, the detailed steps are:

[0068] (1) Primer synthesis: According to the sequence information in Table 1, entrust a relevant biotechnology company to synthesize primers JYSNP-1, JYSNP-2, and JYSNP-3 and choose one of them (only primer JYSNP-2 was synthesized in this example);

[0069] Table 1: Related Primer Sequences

[0070]

[0071] Note: * indicates thio modification.

[0072] (2) Extraction of peach leaf DNA of the population sample to be tested:

[0073] A. Grind about 0.2g of young leaves with liquid nitrogen into powder and place them in a 2mL centrifuge tube;

[0074] B. Add 0.8mL CTAB lysate preheated at 65°C and mix well. The formula of the lysate is: 100mmol / L Tris Cl (pH 8.0), 20mmol / L EDTA (pH 8.0), 1.4mmol / L NaCl, 2.5% CTAB (M / V), 3% PVP (M / V), 2% β-mercaptoethanol (V / V); water bath at 65°C for 1 hour, and shake gently fr...

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Abstract

The invention discloses a molecular marker primer composition capable of identifying freestone / clingstone traits of prunus persica germplasm and application of the molecular marker primer compositionin order to be applied to molecular marker-assisted selection of freestone / clingstone traits of prunus persica. The invention provides the molecular marker primer composition capable of identifying freestone / clingstone traits of prunus persica, which is any one of JYSNP-1, JYSNP-2 and JYSNP-3 and can be used for rapidly indentifying the freestone / clingstone traits of fruits of prunus persica breeding materials. The invention provides a method for assisted screening of excellent prunus persica germplasm resources on the basis of the developed molecular marker primer composition, and the methodhas important guide significance in phenotype prediction of freestone / clingstone traits of the prunus persica and has the advantages of being simple to operate, efficient, rapid, low in cost, accuratein result and the like.

Description

technical field [0001] The invention relates to a group of molecular marker primer combinations and application thereof for identifying the character of peach fruit denuclearization, and belongs to the field of molecular biology. Background technique [0002] Peach [Prunus persica (L.) Batsch] belongs to Rosaceae (Rosaceae), Prunus (Prunus L.), originated in western China, and is one of the important cultivated fruit trees in my country. my country's peach germplasm resources are relatively rich, and using existing resources to breed new and superior varieties with higher economic value is one of the effective ways to improve the quality and yield of my country's peach. Pulp sticking away from the pit is one of the important characteristics of peach fruits; pitting means that when people eat or break the fruit, the pulp can be separated from the peach pit smoothly, and there will not be much pulp left on the peach pit; sticking pit means that the pulp will not It is separat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156C12Q2600/13
Inventor 焦云刘丽琴舒巧云
Owner NINGBO ACAD OF AGRI SCI
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