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A method for identifying female parent of cultivated oat

A technology of oat and female parent, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc. The effect of insufficient evidence

Active Publication Date: 2020-11-10
SHANXI UNIV
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Problems solved by technology

At present, the parents used in the breeding of oat varieties are all determined by screening some morphological characteristics. These morphological characteristics may be caused by the influence of the environment and cannot be inherited in the offspring, thus reducing the breeding efficiency and wasting a lot of money. human and financial resources

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  • A method for identifying female parent of cultivated oat

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Embodiment 1

[0019] Embodiment 1, identification of cultivated oat female parent

[0020] The ccsA-ndhD determination was performed on 667 cultivated oat samples, and the determination steps of each sample were as follows:

[0021] 1. Using the standard CTAB method to extract the total DNA from the cultivated oat leaves;

[0022] 2. Perform PCR amplification on the total DNA extracted in step 1: primer pair for PCR amplification:

[0023] Forward primer: 5'-GGTTTGCATAGTTATGGTTCGT-3' (SEQ ID No.1);

[0024] Reverse primer: 5'-CTGGACCACGGGAACTCTTT-3' (SEQ ID No.2).

[0025] PCR amplification system: double distilled water 23.1ul; 10×tris-HCl buffer (pH8.3, 500mM KCl, 15mMMgCl 2 ) 3ul; dNTP (2.5mM) 0.7ul; SEQ ID No.1 (10uM) 1ul; SEQ ID No.2 (10uM) 1ul; Taq enzyme (5U / ul) 0.2ul; DNA template 1ul (30ng).

[0026] PCR amplification program: 94°C for 5 minutes; then cycle 35 times at 94°C for 10 seconds, 60°C for 20 seconds, and 72°C for 30 seconds; finally incubate at 72°C for 5 minutes.

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Abstract

The invention provides a method for authenticating a cultivating oat female parent. The method comprises the following steps: (1) extracting total DNA of to-be-cultivated oat; (2) performing PCR amplification by taking the total DNA extracted in the step (1) as a template, wherein the sequence of a primer pair of the PCR amplification is shown in SEQ ID No. 1 and SEQ ID No. 2; (3) identifying the female parent of the cultivating oat through the nucleotide sequence the PCR amplification product. The invention further provides a special primer pair for identifying the cultivating oat female parent as well as a kit containing the special primer pair. The cultivating oat is identified by utilizing a molecule biology method, so the problem of insufficient evidence existing in identification by a morphology method is solved, and reasonable and accurate identification of genetic resources of the cultivating oat is realized.

Description

technical field [0001] The invention relates to a method for identifying plant varieties, in particular to a method for identifying the female parent of cultivated oats. Background technique [0002] Oatmeal is an ancient food and grass crop in the world, and it is also an exclusive food for diabetics. Because it is rich in a kind of water-soluble dietary fiber (β-glucan), it has a significant effect on the prevention and treatment of human "three highs", and is popular all over the world. At present, there are hundreds of oat cultivars around the world; in my country, there are also hundreds of oat cultivars that have been bred since the 1960s and 1970s. However, the quality and yield of cultivated oats have not been improved due to the increase of varieties. Therefore, identifying the parents of cultivated oats, clarifying the genetic relationship between oat cultivars, and improving the efficiency of oat breeding have always been the content of people's attention. At p...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895
Inventor 刘兵兵段娜张俊峰
Owner SHANXI UNIV
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