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A method and application of developing specific molecular markers of wheat relative species

A molecular marker and kinship technology, applied in the application field of marker primers and methods, can solve problems such as screening research on specific molecular markers that have not been found in Triticum aestivum

Active Publication Date: 2017-03-08
NANJING AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, after searching, there was no screening research on the specific molecular markers of wheat relative species A. bisativa through high-throughput sequencing.

Method used

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  • A method and application of developing specific molecular markers of wheat relative species
  • A method and application of developing specific molecular markers of wheat relative species
  • A method and application of developing specific molecular markers of wheat relative species

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Embodiment 1

[0188] Choose the Elytrigia in the peak tillering stage, transfer it to a flowerpot filled with quartz sand, culture it in Hogland nutrient solution for two weeks, and transfer it to 250mmol.L -1 Stress was carried out in NaCl solution, and the leaves were cut after 3 hours and 12 hours of stress. Trizol reagent was used to extract the total RNA of Elytrigia sibiricum, and then the samples were tested using the Agilent 2100 (Agilent 2100) platform, and the qualified samples (RIN> 7.5, the total amount is greater than 15ug total RNA) to construct a transcriptome library. Before sequencing on the computer, the qPCR method was further used to check the quality of the transcriptome library to confirm that the fragment size in the library was 330bp and the library concentration was 55.51nM. Finally, 4G-depth de novo sequencing was performed on the Hiseq2500 high-throughput sequencing platform.

[0189] The original sequencing data obtained on the Hiseq2500 platform has been removed rea...

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Abstract

The invention belongs to the technical fields of crop genetics and breeding and agricultural biology and provides marker primers and a method for developing wheat closely related species thinopyrum bessarabicum-specific molecular markers, as well as application of the marker primers and the method. According to the method provided by the invention, 157 primer pairs are provided in total, and thinopyrum bessarabicum-specific fragments after amplification by the primer pairs are totally positioned on thinopyrum bessarabicum chromosomes, indicating the specificity of the primer pairs; simultaneously, the polymorphism rate of the specific amplified fragments obtained by the method for developing the thinopyrum bessarabicum-specific molecular markers, provided by the invention, in thinopyrum bessarabicum is as high as 40.2%, indicating the feasibility of the method. According to the method provided by the invention, 182 molecular markers positioned on the thinopyrum bessarabicum chromosomes are obtained through the 157 primer pairs, and the number is large, thereby providing convenience for excavating thinopyrum bessarabicum beneficial trait genes and further selectively transferring favorable genes into cultivated wheat under assistance of the molecular markers.

Description

Technical field [0001] The invention belongs to the technical field of crop genetic breeding and agricultural biotechnology, and relates to a marker primer and method for developing specific molecular markers of wheat relative species Besamegrass, and applications of the marker primer and method. Background technique [0002] There are abundant beneficial genes in wheat relative species, which are important genetic resources for wheat genetic improvement. Distant hybridization and chromosome engineering techniques are the main ways to transfer and utilize favorable genes from wheat relatives. The development of specific molecular markers for wheat relatives is helpful to accelerate the transfer and utilization of beneficial genes in foreign chromosomes to wheat. [0003] Molecular markers based on PCR (polymerase chain reaction) are simple, effective and economical, and are widely used in molecular marker-assisted selection. At present, the development of PCR-based chromosome-spe...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68C12N15/11C12N15/10
CPCC12Q1/6895C12Q2600/156
Inventor 亓增军李晨旭刘志涛庄丽芳
Owner NANJING AGRICULTURAL UNIVERSITY