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Development and Application of Specific Probes for Tandem Repeat Sequences of Decaploid Echinopsis elongatum

A technology for S. longarum and sequence, which is applied in the field of crop genetics and breeding, can solve the problems of lack of complete genome sequence information, hindering the development and application of genetic resources of D. longarum, and inability to obtain chromosomal-specific molecular markers in batches, etc.

Active Publication Date: 2020-04-07
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] However, with the rapid development of molecular biology, the development of molecular markers of E. elongatum is relatively lagging behind, and the sources of markers are mainly concentrated in its diploid species.
Due to the large genome of the decaploid E. elongatum, the high degree of homology between its genome and the wheat genome, and the lack of complete genome sequence information, the development and research of molecular markers of the decaploid E. elongatum are time-consuming, costly, and expensive. The disadvantage of low efficiency and the inability to obtain chromosome-specific molecular markers in batches have seriously hindered the development and application of E. elongatum genetic resources

Method used

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  • Development and Application of Specific Probes for Tandem Repeat Sequences of Decaploid Echinopsis elongatum
  • Development and Application of Specific Probes for Tandem Repeat Sequences of Decaploid Echinopsis elongatum
  • Development and Application of Specific Probes for Tandem Repeat Sequences of Decaploid Echinopsis elongatum

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Embodiment 1、 10

[0065] Example 1. Development and application of specialized molecular markers and fluorescent probes for decaploid Echinopsis elongatum

[0066] 1. Development and application of specialized molecular markers in decaploid E. elongatum

[0067] 1. Genomic DNA extraction and purification

[0068] Using the CTAB method to extract the genomic DNA of common wheat Chinese spring, decaploid Thipotonia elongatum, octoploid Tripotentia “Xiaoyan 68” and common wheat-decaploid Thipotonia elongatum translocation line, the specific operation steps as follows:

[0069] (1) Take an appropriate amount of tender tissue, freeze it in liquid nitrogen, quickly grind it into powder and put it into a 2mL centrifuge tube.

[0070] (2) Add 800 μL of CTAB extract preheated at 65°C, shake vigorously, place in a water bath at 65°C for 40-60 minutes, and shake gently occasionally.

[0071] (3) Take out the centrifuge tubes, add 800 μL of chloroform:isoamyl alcohol (24:1) to each tube, mix well by inver...

Embodiment 2、 10

[0093] Example 2, Development and Application of Specific Probes for Decaploid Echinopsis elongatum Tandem Repeat Sequence

[0094] Using the decaploid Thiopyrum elongatum (Th. ponticum, 2n=10x=70) as a template, the 67 specialized molecular markers obtained in Example 1 were used for PCR amplification to obtain PCR products. The obtained PCR products were purified and labeled as FISH probes, and in situ hybridization was carried out on the mitotic metaphases of the root tip somatic cells of the octoploid Tritiprum "Xiaoyan 68". Probes with specific hybridization signals on the chromosomes of the decaploid E. elongatum but no hybridization signals on the wheat chromosomes were selected as specialized fluorescent probes for the decaploid E. elongatum. At the same time, the GISH analysis using the whole genome DNA of decaploid E. elongatum as a probe was used as a control, and the specific steps of GISH analysis using the whole genome DNA of decaploid E. elongatum as probes refe...

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Abstract

The invention discloses development and application of a decaploid agropyron elongatum tandem repeat sequence specific probe. According to the development and the application, a specific length amplified fragment sequencing technology (SLAF-seq) is utilized to perform sequencing on common wheat, a common wheat-decaploid agropyron elongatum translocation line and decaploid agropyron elongatum, a decaploid agropyron elongatum specific sequence is acquired through sequence data comparative analysis, and the tandem repeat sequence specific probe is developed based on the decaploid agropyron elongatum specific sequence. The tandem repeat sequence specific probe provided by the invention can be used for rapidly identifying decaploid agropyron elongatum chromosomes and identifying a translocation line in a process that chromosome fragments of agropyron elongatum are transferred into the wheat, providing an important way of molecular marker-assisted selection to improve the wheat hereditary property and also providing an important application basis for wheat molecular breeding, phyletic evolution and germplasm resource identification.

Description

technical field [0001] The invention belongs to the field of crop genetics and breeding, and in particular relates to the development and application of specific probes for the tandem repeat sequence of the decaploid Etyophony elongatum. Background technique [0002] Wheat (Triticum aestivm) is the second largest food crop with the largest planting area in the world and its output is second only to corn. In the past half century, the total output of wheat in the world has more than tripled, and excellent wheat varieties have played a decisive role in increasing wheat output. There are a large number of beneficial genetic resources that wheat lacks in wild relatives of wheat, and fully developing and utilizing these excellent genetic resources to breed new wheat varieties has become one of the goals of wheat breeding. Thinopyrum ponticum (Thinopyrum ponticum, 2n=10x=70) is a perennial herb belonging to the genus Thinopyrum ponticum in the Poaceae family, and is an important ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6869C12Q1/6895C12Q2600/13C12Q2535/122C12Q2525/151C12Q2563/107
Inventor 郑琪刘利勤滕婉罗巧玲李滨李宏伟李振声
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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