Method for rapidly identifying switchgrass hybrids by SSR (simple sequence repeat) molecular marker technology

A molecular marker, hybrid technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve problems such as high cost and time-consuming, and achieve high accuracy, easy operation, and good versatility.

Inactive Publication Date: 2012-10-03
SICHUAN AGRI UNIV
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Takes more time and costs more

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  • Method for rapidly identifying switchgrass hybrids by SSR (simple sequence repeat) molecular marker technology
  • Method for rapidly identifying switchgrass hybrids by SSR (simple sequence repeat) molecular marker technology
  • Method for rapidly identifying switchgrass hybrids by SSR (simple sequence repeat) molecular marker technology

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Embodiment Construction

[0022] The present invention will be described in detail below in conjunction with specific embodiments.

[0023] 1.2.1 Test materials: hybrid offspring of switchgrass to be identified and their parent plants or seeds.

[0024] 1.2.2 Genomic DNA extraction

[0025] Using the young leaves or seeds of switchgrass plants to be identified as materials, DNA was extracted using a plant genomic DNA extraction kit (Tiangen Company). The purity of the extracted DNA was tested by 0.8% agarose gel electrophoresis. DNA samples were stored in a -20°C refrigerator. When the experiment was started, a part of each DNA sample was taken out and diluted to 20 ng / μL, and stored in a refrigerator at 4°C for use.

[0026] 1.2.3 SSR Analysis

[0027] Using the screened switchgrass EST-SSR marker primers:

[0028] primer1-F: 5'-AGTTTGAGCTTTGTGTTTTTG-3',

[0029] primer1-R: 5'-CAACAATTTGGTTTCAACAAG-3';

[0030] Perform hybrid identification. The sequence has the characteristics of clear and st...

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Abstract

The invention discloses a method for rapidly identifying switchgrass hybrids by an SSR (simple sequence repeat) molecular marker technology. The method comprises the following steps of: A1, extracting genomic DNA (deoxyribonucleic acid); A2, performing SSR analysis; and A3, identifying the hybrids. Primers are designed by means of EST (expressed sequence tag) sequences, SSR primers which are obtained by development and have high polymorphism, stable and clear banding patterns and parents complementary bands are used for performing PCR (polymerase chain reaction) amplification on DNA of plants or seeds of the switchgrass hybrids, amplification products are separated by means of agarose gel electrophoresis, and the hybrids with male parent specificity bands are genuine hybrids. The method has the advantages of convenience for operation, low cost, high accuracy and high versatility, and can be used for rapidly identifying the genuineness of the hybrids combined by different parents of switchgrass.

Description

technical field [0001] The invention relates to a method for rapidly identifying hybrid species of switchgrass by using SSR molecular marker technology. Background technique [0002] Switchgrass (Panicum virgatum) is a plant of the genus Panicum in the Gramineae family. It has tall plants, good adaptability, tolerance to barrenness, and environmental friendliness. It is now regarded as the first choice among cellulosic energy plants. But its breeding level is backward, and molecular breeding can greatly speed up its breeding process. Hybrid identification is the basic work of genetic breeding, and the identified true hybrids can provide an important basis for the construction of genetic maps. Switchgrass is a cross-pollinated plant with a certain self-fertilization rate. How to quickly and accurately identify the authenticity of hybrid offspring of switchgrass is the basic work of genetic breeding. [0003] In addition to morphological identification, molecular marker tech...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
Inventor 黄琳凯张新全张婧曾兵
Owner SICHUAN AGRI UNIV
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