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Green smut bug real-time fluorescent quantitative PCR test kit and its use

A fluorescence quantitative, rice smut fungus technology, applied in the direction of microbial determination/inspection, fluorescence/phosphorescence, biochemical equipment and methods, etc., can solve the problem of inability to calculate the initial DNA copy number, etc. High specificity and good reproducibility

Inactive Publication Date: 2006-10-25
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this traditional PCR method measures the end product of PCR, not the copy number of the starting DNA
Since there is no linear relationship between the final PCR product and the initial template amount, the initial DNA copy number cannot be calculated from the final PCR product amount

Method used

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  • Green smut bug real-time fluorescent quantitative PCR test kit and its use
  • Green smut bug real-time fluorescent quantitative PCR test kit and its use
  • Green smut bug real-time fluorescent quantitative PCR test kit and its use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1: detection of rice smut bacteria amount on field duckweed in rice growing season

[0031] In the experimental base of the Chinese Academy of Agricultural Sciences in Changping District, Beijing, two paddy fields were selected: one was a field where the serious rice smut disease occurred in 2004, and the other was a field that had been planted with soybeans before 2004 and planted rice in the first year of 2004, located downstream of the severely diseased paddy field The rice varieties planted in the two fields are the rice smut-susceptible variety Quality No. 1, and the rice seedling raising and transplanting periods are the same. On July 15, 2005, duckweed was obtained from the water surface of the above two paddy fields, and the duckweed retrieved from the field was immediately freeze-dried. Take 5 mg of freeze-dried standard sample and adopt MasterPure from Epicentre Company TM DNA Purification Kit extracts and purifies DNA. The DNA was dissolved in 2...

Embodiment 2

[0036] Example 2: Detection of rice smut bacteria inside the spikelets of plants in the reproductive growth period of rice

[0037] In early August 2004, samples were taken from the paddy fields of Wangjia Township, Dongling District, Shenyang during the reproductive growth period of rice, and the samples were young panicles at the opening stage. The sample was cleaned repeatedly, and after microscopic examination confirmed that the surface of the sample was free of hyphae and spores, Epicentre's MasterPure was used to TM DNA Purification Kit extracts and purifies DNA. The DNA of each young ear was dissolved in 20 μl TE buffer, and 1 μl of the DNA solution was used as a template, and the primer pairs US3-3 / US4-5 (first round) and US1-3 / US2-5 (second round) were used for each sample Nested PCR (nested PCR) detection was carried out, and the second round of PCR used 1 μl of the first round of PCR product as a template. The amplified products were detected by 1.2% agarose elec...

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Abstract

The invention relates to a green smut germ real time fluorescence fix quantity PCR agent box and the application. The box uses Taqman and positive primer, negative primer and masculine standard product to build up real time fluorescence PCR reacting system that could rapidly, and quantified testing green smut germ. It is suited for farm environment. It is also could be used to testing the germ testing for paddy seed and the multiplication speed in different paddy kinds.

Description

technical field [0001] The invention relates to a real-time fluorescence quantitative PCR kit for detecting rice smut bacteria, which comprises rice smut bacteria-specific primers and matched fluorescent labeling probes, and is suitable for detecting rice smut bacteria in farmland environments (including the amount of overwintering bacteria in soil and paddy fields) It can also be used for real-time monitoring of rice smut bacteria on duckweed and rice plants; it can also be used for the detection of rice seed bacteria and the comparison of the proliferation speed of different strains of smut bacteria in different rice varieties. Background technique [0002] Rice false smut caused by Ustilaginoidea virens (Cooke) Tak. is a worldwide rice fungal disease, which is serious in China, Japan, India and other countries. Rice false smut is a disease of the ear of rice. The pathogen turns the grain into a rice ball, covered with a large number of powdery, dark green chlamydospores. ...

Claims

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Application Information

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IPC IPC(8): G01N21/64C12Q1/68
Inventor 周永力谢学文徐建龙王疏黎志康
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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