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Bemisia tabaci(Gennadius) biological B type and Q type specific primers and quick identification method

A technology of specific primers and identification methods, applied in the fields of identification and control, biological type B and Q type specific primers and rapid identification of Bemisia tabaci, and detection of crop pests

Inactive Publication Date: 2012-04-11
INST OF PLANT PROTECTION FAAS
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  • Summary
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The purpose of the present invention is to provide a method for the detection and identification of Bemisia tabaci biotype in the prior art, which requires a long cycle, poor specificity and low sensitivity. Specific primers for biotype B and Q of Bemisia tabaci, and a rapid identification method with reliable results, easy operation, strong specificity and high sensitivity

Method used

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  • Bemisia tabaci(Gennadius) biological B type and Q type specific primers and quick identification method
  • Bemisia tabaci(Gennadius) biological B type and Q type specific primers and quick identification method

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Effect test

Embodiment 1

[0037] From the adult samples of Bemisia tabaci collected in the field, the SDS method was used to extract DNA from a single head. The specific process was as follows: a. Female individuals were screened from each adult Bemisia tabaci sample; b. Single female adults were placed in PCR centrifugation Add 30μl lysate solution (1%SDS, 25mmol / L Nacl, 25mmol / L EDTA, 10mmol / L Tris-Hcl pH8.0), grind thoroughly with a grinding rod, then add 10μl proteinase K, and mix well; c. Water bath at 55°C for 1 hour, shake and mix occasionally; d. Add 40 μl chloroform and mix (note that the mixing should not be too vigorous to ensure the integrity of the DNA), 10,000 rpmCentrifuge for 5 minutes; e. Take 30ul of the supernatant, add it to a PCR centrifuge tube, add 90μl of ice-prepared absolute ethanol, and place it at -20°C for 10min. 10,000 rpm Centrifuge for 5 minutes to remove ethanol; f, add 100 ul of 70% ethanol to wash once, invert and dry at room temperature for 10 minutes; g, dissolve...

Embodiment 2

[0045] The PCR reaction system is as in Example 1.

[0046] 3. Test result: if figure 2 As shown, in the 20μl reaction system, eight samples of Bemisia tabaci genomic DNA can obtain obvious amplification target bands except the first sample without obvious amplification target bands, the product size is 300pb, and the detection sensitivity can reach 1ng.

[0047] Institute of Plant Protection, Fujian Academy of Agricultural Sciences

[0048]

[0049] Biological type B and Q specific primers and rapid identification method of Bemisia tabaci

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Abstract

The invention discloses bemisia tabaci(Gennadius) biological B type and Q type specific primers and a quick identification method. According to the invention, two pairs of specific primers (B type 5'-CTAGGGTTTATTGTTTGAGGTCATCATATATTC-3'; 5'-AATATCGACGAGGATTCCCCCT-3') and (Q type 5'-CTTGGTAACTCTTCTGTAGATGTGTGTT-3'; 5'-CCTTCCCGCAGAAGAAATTTTGTTC-3') are respectively designed aiming at bemisia tabaci(Gennadius) biological B type and Q type, and a double primer PCR(Polymerase Chain Reaction) amplification system suitable for quick identification of bemisia tabaci(Gennadius) biological types is established. The method can be applied to quick identification and distribution situation investigation of alien invasive bemisia tabaci(Gennadius) biological types in the fields, and provides a scientific basis for establishment of control strategies.

Description

technical field [0001] The present invention relates to a kind of specific primers and rapid identification method of Bemisia tabaci biological type B and Q type, which is specially used for rapid, sensitive and accurate molecular identification of alien invasive biological species B. tabaci and Q type, and can be used for field tobacco powder at the same time Monitoring and identification of lice colony biotypes. The invention belongs to the field of crop pest detection, identification and control technology. Background technique [0002] whitefly Bemisia tabaci (Gennadius) is widely distributed in all continents except Antarctica, and is an important pest in tropical, subtropical and adjacent temperate regions (Brown et al., 1995). Bemisia tabaci is just a compound group in a continuous evolution process, with multiple biotypes. Bemisia tabaci can be divided into different biotypes according to the biological differences of different geographical populations in host ra...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 何玉仙郑宇赵建伟栗丽娜翁启勇
Owner INST OF PLANT PROTECTION FAAS
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