Multiple PCR (Polymerase Chain Reaction) kit for quickly identifying plant hopper species and identifying method thereof

A kit and planthopper technology, applied in the field of molecular biology identification, can solve the problems of high cost and long time, and achieve the effects of saving experimental time, easy operation and high repeatability

Inactive Publication Date: 2018-02-02
CHINA NAT RICE RES INST
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Cui Yali et al. (2012) used the ITS region to design primers and used DNA as a template to initially establish a multiplex PCR detection technology for brown planthopper, brown planthopper and pseudo brown planthopper, but it was time-consuming and costly. There is a certain difference in the length of the amplified band

Method used

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  • Multiple PCR (Polymerase Chain Reaction) kit for quickly identifying plant hopper species and identifying method thereof
  • Multiple PCR (Polymerase Chain Reaction) kit for quickly identifying plant hopper species and identifying method thereof
  • Multiple PCR (Polymerase Chain Reaction) kit for quickly identifying plant hopper species and identifying method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] Materials and Methods

[0040] 1. Materials

[0041] 1.1 Samples of brown planthopper, brown planthopper, pseudo brown planthopper, white-backed planthopper and brown planthopper from Zhaoping, Guangxi, Wan'an, Jiangxi and Fuyang, Zhejiang:

[0042] 1.2 Main instrument reagents

[0043]

[0044] 2. Method

[0045] 2.1 Obtaining crude tissue fluid

[0046] a. Take a single planthopper to be tested and put it into a 2 ml centrifuge tube;

[0047] b. Add 200 ml of distilled water, grind with an automatic grinder, 8000 rpm × 1 min, and grind 384 samples at a time;

[0048] c. The grinding liquid is the crude tissue fluid of planthoppers without centrifugation.

[0049] 2.2 PCR reaction

[0050] PCR amplification system:

[0051]

[0052] PCR reaction conditions:

[0053]

[0054] 2.0% agarose gel electrophoresis detection.

[0055] Take 8 μL of the amplified product and spot on 2.0 % (W / V) agarose gel, which contains 0.1-0.5 g / ml of Gelred, with 100 bp Mar...

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Abstract

The invention relates to a multiple PCR (Polymerase Chain Reaction) kit for quickly identifying plant hopper species and an identifying method thereof, which belong to the technical field of molecularbiology identification. The kit comprises (1) three upstream primers and three downstream primers; (2) a positive control recombinant plasmid template; (3) a PCR reaction fluid. According to the multiple PCR kit for quickly identifying the plant hopper species and the identifying method thereof, on the basis of aiming at an ITS sequence of three plant hoppers and verifying the ITS area sequence,a primer pair and a matching ratio are designed and optimized, and a coarse tissure sample is used as a template, so that the time and the reagent consumed for extracting DNA are saved, the multiple PCR kit for quickly, accurately, sensitively, high-throughput and low-cost identifying the three plant hoppers and the identifying method are built, and options are provided for identifying brown planthoppers, imitated brown planthoppers and fake brown planthoppers through a molecular biology method.

Description

technical field [0001] The invention belongs to the technical field of molecular biology identification, and in particular relates to a multiplex PCR kit for quickly identifying planthopper species and an identification method thereof. Background technique [0002] Brown planthopper[ Nilaparvata lugens (Stål)] is one of the most serious rice pests in Asian rice regions, with long-distance migratory habits. Accurate forecasting of pest situation is the key to pest control. At present, lamp lures are mainly used for monitoring. The date of first sighting, the amount of migration and the peak of migration of N. lugens under the forecasting light are the important basis for the prediction of N. lugens and the key information basis for the formulation of the control strategy of N. lugens. [0003] Nilaparvata lugens[ Nilaparvata bakeri (Muir)] and pseudo brown planthopper [Nilaparvata muiriChina] are similar species of the same genus of brown planthopper, which do not use ri...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/686C12N15/11
CPCC12Q1/686C12Q1/6888C12Q2600/16C12Q2537/143
Inventor 刘淑华唐健刘瑞罗举王爱英杨保军
Owner CHINA NAT RICE RES INST
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