Molecular marker for rapid detection of high-yield gene of Thinopyrum elongatum and its application

A technology of wheat grass and molecular markers, which is applied in the field of molecular genetics and breeding, can solve the problems of lack of high-yield gene molecular markers, time-consuming, and difficulty in phenotypic identification.

Active Publication Date: 2019-01-25
CROP RES INST SHANDONG ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, traditional breeding methods are time-consuming, laborious, difficult to identify phenotypes, and have low breeding efficiency. Molecular mark

Method used

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  • Molecular marker for rapid detection of high-yield gene of Thinopyrum elongatum and its application
  • Molecular marker for rapid detection of high-yield gene of Thinopyrum elongatum and its application
  • Molecular marker for rapid detection of high-yield gene of Thinopyrum elongatum and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0059] Example 1 Creation of Near-Isogenic Lines of High-yielding Genes of Wheat-Thinopyrum elongatum

[0060] In 2015, the hybrid combination (Liangxing 66 × SNK064) was configured in the greenhouse of the Crop Research Institute of Shandong Academy of Agricultural Sciences, and obtained F 1 ; Spring 2016, will receive F 1 Backcross with Liangxing 66 and get BC 1 f 1 , and carry out molecular marker identification on the obtained plants, on the basis of molecular marker identification, the BC carrying target gene and exogenous fragment 1 f 1 Continue to backcross with Liangxing 66 and get BC 2 f 1 . In the summer and autumn of 2016, the artificial climate chamber continued to add generations, self-crossed twice, and obtained BC 2 f 3 , with the help of molecular markers and fluorescence in situ hybridization, a wheat-Thinopyrum elongatum 7el2L heterozygous line was obtained; in the winter of 2016 and the spring of 2017, the above heterozygous line continued to be self...

Embodiment 2

[0062] Example 2 Analysis of agronomic and yield-related traits of high-yielding near-isogenic lines of Echinopsis elongatum

[0063] For in situ hybridization using Oligo-pTa535.1 and Oligo-pSc119.2 as probes, the probe synthesis, probe concentration and specific operation steps refer to the method of Tang et al. [2014]. After the in situ hybridization chromosome preparation was dried naturally, 0.25 μg / mL propidium iodide was added dropwise and covered with a cover slip, and photographed under an Olympus BX51 fluorescence microscope.

[0064] The above-mentioned near-isogenic line materials (SAAS1001, SAAS1002 and SAAS1003) were planted in the Lingcheng Experimental Base in the winter of 2017. The area of ​​the plot was 1.5 m × 6.0 m, with 3 replicates. For the flowering stage and grain filling stage of wheat, the length and width of the flag leaf, the length of the ear, the number of grains per ear, the number of fertile spikelets, the number of sterile spikelets, etc. 6 t...

Embodiment 3

[0066] Example 3 Development of Tightly Linked Molecular Markers for High Yield Genes of Echinopsis elongatum

[0067] According to the sequence information of the published wheat genome (7DL end), SSR primers were designed using Primer 3.0, and wheat varieties Jimai 22, Liangxing 66, SAAS1001 and Jimai 22 mixed DNA, SAAS1002 and Liangxing 66 mixed DN, SAAS1003 and SAAS1001 Mixed DNA and wheat-Thinopyrum elongated near-isogenic lines SAAS1001, SAAS1002 and SAAS1003 were used as materials for PCR amplification to search for molecular markers closely linked to high-yielding genes.

[0068] In the PCR amplification reaction system, the volume of the PCR reaction system is 10.0 μL, including 10 × Buffer buffer: 1.0 μL, 2.5 mM dNTP solution: 0.8 μL, Taq enzyme 0.1 μL, sterile double distilled water: 4.1 μL, 50 ng DNA template 2.0 μL (Jimai 22, Liangxing 66, SAAS1001 and Jimai 22 mixed DNA, SAAS1002 and Liangxing 66 mixed DNA, SAAS1003 and SAAS1001 mixed DNA, and wheat-Ethiopia elon...

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Abstract

The invention belongs to the field of genetics, a molecular marker for rapid detection of high yield gene of Thinopyrum elongatum and its application are provided, by conventional hybridization, backcrossing and greenhouse and artificial climate chamber generation techniques, a near-isogenic line of high yield gene 7el2L of Thinopyrum elongatum was created. Combined with fluorescence in situ hybridization (FISH) and analysis of agronomic and yield-related traits, molecular markers closely linked to 7el2L were obtained, which could be used in wheat breeding for high yield and early molecular-assisted selection for yield-related traits, in order to improve breeding efficiency.

Description

technical field [0001] The invention belongs to the field of molecular genetic breeding, and in particular relates to a molecular marker for rapidly detecting the high-yield gene of Echinopsis elongatum and its application. Background technique [0002] Wheat (Triticum aestivum L.) is one of the most important food crops in the world, its yield is second only to corn and rice, and it is the third largest food crop. The world's wheat planting area accounts for 17.0% of the world's total grain planting area (217 million hectares), and my country's annual wheat planting area is about 25 million hectares, accounting for 21.8% of the country's total grain planting area (113 million hectares) (FAO, 2012 ; National Bureau of Statistics of China, 2014). Wheat is one of the most important food crops because it feeds 40% of the world's population and provides 20% of human energy and protein supply. According to reports, by 2050, the total population of the world will reach more than ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 郭军刘建军李豪圣宋健民刘成曹新有刘爱峰赵振东
Owner CROP RES INST SHANDONG ACAD OF AGRI SCI
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