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Primer group, kit and application of primer group in rapid identification of pepper root rot bacteria

The technology of a primer set and a kit, which is applied in the application field of primer sets and rapid identification of pepper root rot bacteria, can solve the problems of inaccurate identification results and complicated operation methods, shorten the time required for identification, simplify the identification process, Improve the effect of anti-efficacy

Inactive Publication Date: 2022-02-01
INST OF PLANT PROTECTION SICHUAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the wide variety of Fusarium genus, some Fusarium species have multiple specialized types, and the morphological characteristics between species are relatively close. The identification of Fusarium solani only by morphological characteristics is not only complicated in operation, but also inaccurate in identification results.

Method used

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  • Primer group, kit and application of primer group in rapid identification of pepper root rot bacteria
  • Primer group, kit and application of primer group in rapid identification of pepper root rot bacteria
  • Primer group, kit and application of primer group in rapid identification of pepper root rot bacteria

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] A method for rapid identification of prickly ash root rot bacteria (fusarium solani), comprising the following steps:

[0036] S1. Synthetic primer set: the primer set includes upstream primer F1 and downstream primer R1; the sequence of upstream primer F1 is: 5'-CGTGTCGCAGAAGAGGG-3', and the sequence of downstream primer R1 is: 5'-AATAGGAAGCCGCTGAGC-3'; The synthetic concentrations of primer F1 and downstream primer R1 were both 10 μmol / L;

[0037] S2. Extract sample DNA: Using conventional DNA extraction means (CTAB method), extract the concentration of 50ng / μL from Fusarium solani, Fusarium oxysporum, Fusarium graminearum, Fusarium moniliforme and Botrytis cinerea respectively. DNA diluent to obtain multiple sets of sample DNA;

[0038] S3. Prepare the PCR reaction system: add the sample DNA of each group into their respective 25 μL reaction systems, and each 25 μL reaction system includes the following components:

[0039]

[0040]

[0041] S4. Routine PCR d...

Embodiment 2

[0046] A method for rapid identification of prickly ash root rot bacteria (fusarium solani), comprising the following steps:

[0047] S1. Synthetic primer set: the primer set includes upstream primer F1 and downstream primer R1; the sequence of upstream primer F1 is: 5'-CGTGTCGCAGAAGAGGG-3', and the sequence of downstream primer R1 is: 5'-AATAGGAAGCCGCTGAGC-3'; The synthetic concentrations of primer F1 and downstream primer R1 were both 10 μmol / L;

[0048] S2. Extracting sample DNA: using conventional DNA extraction means (CTAB method), extracting a DNA dilution with a concentration of 50 ng / μL from the rhizosphere soil of the pathogenic Zanthoxylum bungeanum root inoculated with Fusarium solani to obtain sample DNA;

[0049] S3. Prepare the PCR reaction system: add the sample DNA into the 25 μL reaction system, which includes the following components:

[0050]

[0051] S4. Routine PCR detection: According to the above-mentioned PCR reaction system, use the sample DNA as a t...

Embodiment 3

[0056] A method for rapid identification of prickly ash root rot bacteria (fusarium solani), comprising the following steps:

[0057] S1. Synthetic primer set: the primer set includes upstream primer F1 and downstream primer R1; the sequence of upstream primer F1 is: 5'-CGTGTCGCAGAAGAGGG-3', and the sequence of downstream primer R1 is: 5'-AATAGGAAGCCGCTGAGC-3'; The synthetic concentrations of primer F1 and downstream primer R1 were both 10 μmol / L;

[0058] S2. Extract sample DNA: Using conventional DNA extraction means (CTAB method), extract the concentration of 50ng / μL from the healthy Zanthoxylum bungeanum roots that have not been inoculated with Fusarium solani and the diseased Zanthoxylum bungeanum roots that have been inoculated with Fusarium solani, respectively. DNA diluent to obtain 2 sets of sample DNA;

[0059] S3. Prepare the PCR reaction system: Add the two groups of sample DNA into their respective 25 μL reaction systems, and the 25 μL reaction systems of each gr...

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Abstract

The invention relates to the technical field of molecular detection, and discloses a primer group and a kit containing the primer group. The primer group comprises an upstream primer F1 and a downstream primer R1, and the sequences of the upstream primer F1 and the downstream primer R1 are as follows: the upstream primer F1 is 5'-CGTGTCGCAGAAGAGGG-3'; the downstream primer R1 is 5'-AATAGGAAGCCGCTGAGC-3', and the invention also discloses an application of the primer group or the kit in rapid identification of pepper root rot bacteria. The application comprises the following steps: S1, extracting sample DNA; S2, taking the sample DNA as a template, and carrying out PCR amplification reaction by utilizing the primer group to obtain a PCR amplification product; and S3, analyzing a PCR amplification product. The primer group provided by the invention can specifically identify fusarium solani, and achieves the characteristics of high amplification efficiency, strong specificity and high accuracy; meanwhile, the rapid identification method established in combination with the primer group is quite simple, and fusarium solani can be identified without morphological identification and DNA sequencing.

Description

technical field [0001] The invention relates to the technical field of molecular detection, in particular to a primer set, a kit and their application in rapid identification of pepper root rot bacteria. Background technique [0002] Zanthoxylum bungeanum Maxim is a small deciduous tree belonging to Rutaceae and the genus Zanthoxylum. It is drought-tolerant and sunshine-loving, and is widely planted in various places. The peel of Zanthoxylum bungeanum can be used as seasoning, extract aromatic oil, and be used as medicine. The seeds are edible, and can also be processed to make soap. [0003] Root rot is a disease caused by a fungus. This disease will cause the roots to rot, the function of absorbing water and nutrients will gradually weaken, and finally the whole plant will die, mainly manifested as yellowing and withering of the leaves of the whole plant. Pepper root rot is mainly a soil-borne disease caused by Fusarium solani. It often occurs in nurseries and adult peppe...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12Q1/686C12Q1/04C12N15/11C12R1/77
CPCC12Q1/6895C12Q1/686C12Q2565/125
Inventor 杨潇湘刘勇张蕾黄小琴张重梅唐蓉鲜赟曦周西全
Owner INST OF PLANT PROTECTION SICHUAN ACAD OF AGRI SCI
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