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Method for developing wheat closely related species-specific molecular markers and application thereof

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: 2015-01-07
NANJING AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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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  • Method for developing wheat closely related species-specific molecular markers and application thereof
  • Method for developing wheat closely related species-specific molecular markers and application thereof
  • Method for developing wheat closely related species-specific molecular markers and application thereof

Examples

Experimental program
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Embodiment 1

[0188] Select Thinopyrum baisa in the prime tillering stage, transfer it to a flowerpot filled with quartz sand, cultivate it in Hogland nutrient solution for two weeks, and then transfer it to 250mmol.L -1 The stress was carried out in NaCl solution, and the leaves were clipped after 3 hours and 12 hours of stress. The Trizol reagent was used to extract the total RNA of Thiopyrum basina, and then the Agilent2100 (Agilent 2100) platform was used to detect the samples, and the transcriptome library was constructed for the qualified samples (RIN>7.5, the total amount was greater than 15ug total RNA). Before sequencing on the machine, the quality of the transcriptome library was further checked by qPCR method, and the fragment size in the library was confirmed to be 330bp and the library concentration was 55.51nM. Finally, de novo sequencing was performed at a depth of 4G on the Hiseq2500 high-throughput sequencing platform.

[0189] The raw sequencing data obtained on the Hiseq2...

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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 field of crop genetics and breeding and agricultural biotechnology, and relates to a marker primer and a method for developing specific molecular markers of a wheat relative species A. bisativa, and an application of the marker primer and method. Background technique [0002] Wheat relative species contain abundant beneficial genes, which are important genetic resources for wheat genetic improvement. Distant hybridization and chromosome engineering are the main ways to transfer and utilize favorable genes of wheat relative species. The development of specific molecular markers for wheat relative species can help 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 marker-assisted selection. At present, the development of PCR-based chromosome-specific mo...

Claims

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Application Information

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Patent Type & Authority Applications(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