A kind of primer for identifying Zoysia species and the identification method of Zoysia species

A technology of Zoysia species, which is applied in biochemical equipment and methods, microbiological measurement/inspection, DNA/RNA fragments, etc., can solve the problems that the test results are easily affected by environmental factors and the cycle is long, and achieve the goal of improving grass Variety approval work quality, reliable and quick method, convenient and quick operation effect

Active Publication Date: 2018-02-13
CHINA AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Test results are easily affected by environmental factors, and the cycle is long
And with the increasingly narrow genetic basis of breeding materials and the increasing number of breeding varieties, the differences between varieties are getting smaller and smaller, and it is difficult to accurately identify them solely by traditional botanical characters.

Method used

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  • A kind of primer for identifying Zoysia species and the identification method of Zoysia species
  • A kind of primer for identifying Zoysia species and the identification method of Zoysia species
  • A kind of primer for identifying Zoysia species and the identification method of Zoysia species

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Experimental program
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Effect test

Embodiment 1

[0028] 1) Extraction of Zoysias Genomic DNA:

[0029] Number of samples: For each variety sample, 25 populations are used as a mixed sample.

[0030] 2) Reagent preparation method:

[0031] Formula of DNA extraction solution: Weigh D-sorbitol 6.375g, Tris-base 1.21g, EDTA-Na 2 0.168g, dilute to 100ml with sterile water, adjust the pH range to 7.5-8.25, and store at 4°C.

[0032] Nuclear lysate formula: weigh Tris-base 2.42g, EDTA-Na 2 1.86g, sodium chloride 11.69g, CTAB 2g, dilute to 100ml with sterile water, adjust the pH range to 7.5-8.0, and sterilize under high temperature and high pressure.

[0033] 5% sodium lauryl sarcosinate formula: weigh 5g of sodium lauryl sarcosinate, dilute to 100ml with sterile water, filter with filter paper, and store at room temperature.

[0034] 3) The leaves of Zoysia species were used as raw materials, and the total genome DNA was extracted by CTAB method. The specific operation was as follows: 0.1 g of fresh leaf tissue was weighed, p...

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Abstract

The invention relates to a primer and method for recognizing zoysia japonica varieties. The nucleotide sequences of the primer are shown as the SEQ ID No.1 and the SEQ ID No.2. The invention further provides a method for recognizing the zoysia japonica varieties. The method comprises the steps that a genome DNA of a sample to be tested is used as a template DNA, and PCR amplification is carried out by means of the primer. Compared with an existing three-nature testing and recognizing method (namely the appearance, morphological characteristic description method) for recognizing zoysia japonica varieties, testing results are not affected by environment factors, operation is convenient and rapid, the testing period is obviously shortened, and the testing results are more accurate and reliable.

Description

technical field [0001] The invention belongs to the field of biological identification, and in particular relates to a primer for identifying zoysia varieties and a method for identifying zoysia varieties. Background technique [0002] China is the country with the richest zoysia resources in the world, and at the same time, zoysia is also the only turfgrass species whose seeds are exported in my country. The agricultural industry standard "Technical Regulations for the Approval of Grass Varieties" points out that the bred varieties should have distinctness, uniformity and stability (referred to as "DUS" or "DUS"). Therefore, before the approval of grass varieties, in addition to completing the uniformly arranged regional tests, new grass varieties should also pass the "three characteristics" test. At present, the method of "three characteristics" testing and identification of new grass varieties usually adopts the method of describing appearance and morphology. The test r...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 李曼莉王晨郑丽华何祖欣
Owner CHINA AGRI UNIV
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