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Bemisia tabaci AsiaII3 hidden species transient receptor ion channel gene specific probe and application thereof

An ion channel, technology of Bemisia tabaci, applied in the field of agricultural biology

Inactive Publication Date: 2020-05-08
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The expression of TRP gene in Bemisia tabaci can be induced by temperature stress. For example, the expression of TRP gene in the invasive species MED cryptic species reaches the highest level under the conditions of extreme high temperature 40°C and extreme low temperature 0°C. However, the expression of TRP gene in the native species AsiaII3 cryptic species Currently unreported

Method used

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  • Bemisia tabaci AsiaII3 hidden species transient receptor ion channel gene specific probe and application thereof
  • Bemisia tabaci AsiaII3 hidden species transient receptor ion channel gene specific probe and application thereof
  • Bemisia tabaci AsiaII3 hidden species transient receptor ion channel gene specific probe and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Cloning of full-length cDNA sequence of BtTRP gene of Bemisia tabaci AsiaII3 cryptic species

[0028] Take 200 adults of Bemisia tabaci under different temperature stress conditions and put them into 1.5mL centrifuge tubes, freeze them in liquid nitrogen, grind them into powder with a grinding rod, then extract RNA, and store them at -80°C for later use. Using cDNA as a template, primers were designed for PCR amplification. The designed primers are shown in Table 1 and Table 2:

[0029] Table 1

[0030]

[0031] Table 2

[0032]

[0033] The experiment found that the full-length cDNA of the BtTRP gene of the AsiaII3 cryptic species of Bemisia tabaci was obtained by PCR amplification using the primers in Table 1. However, the target gene sequence could not be obtained using the primers in Table 2.

[0034] Using the sequence in Table 1, through PCR amplification, the cDNA sequence full length of the BtTRP gene obtained is 3821bp, the resulting gene ha...

Embodiment 2

[0035] Example 2 in situ hybridization

[0036] (1) Specific mRNA in situ hybridization probe for BtTRP gene of whitefly

[0037] (a) Using cDNA as a template, design primers to amplify common PCR products. The designed primers are shown in Table 3 and Table 4.

[0038] table 3

[0039]

[0040] Table 4

[0041]

[0042] The experiment found that using the primers in Table 3, the middle fragment of the BtTRP gene of the AsiaII3 cryptic species of Bemisia tabaci was amplified by PCR, but the target gene sequence could not be obtained by using the primers in Table 4.

[0043] The product was recovered and purified by gel cutting, and the purified PCR product was cloned and sequenced using pGEM-T Easy competent cells. The sequencing sequence is compared and analyzed, and the cloned fragment obtained is a fragment of the target gene;

[0044] (b) Pick the positive clones verified by sequencing, operate according to the instructions in the AxyPrep Plasmid DNA Mini Kit, and...

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Abstract

The present invention belongs to the field of agricultural biology and specifically relates to a bemisia tabaci AsiaII3 hidden species transient receptor ion channel gene specific probe and an application thereof. A nucleotide sequence of the bemisia tabaci AsiaII3 hidden species transient receptor ion channel gene specific probe is shown as SEQ No.1. The BtTRP probe sequence has a clear specificity for display of BtTRP gene mRNA and realizes the display function of the BtTRP gene mRNA level. The specific probe is directly used for adult to conduct in situ hybridization, a slice-manufacturingstep is saved, complexity of experiments can be reduced, experiment steps are reduced, and besides, the specific probe has higher sensitivity and can be more convenient and quick to detect localization expression of target genes in insect body.

Description

technical field [0001] The invention belongs to the field of agricultural biology, and in particular relates to a specific probe for a transient sensor ion channel gene of the whitefly AsiaII3 cryptic species and an application thereof. Background technique [0002] Bemisia tabaci (Gennadius), belongs to Hemiptera (Hemiptera), whitefly family (Aleyrodidae), small whitefly belongs to Bemisia, has strong fecundity and wide host range, mainly through direct sucking, secreting honey Many kinds of crops and garden flowers are harmed by dew, spreading plant viruses, etc., causing great economic losses. Bemisia tabaci is a species complex containing more than 36 cryptic species. At present, there are 15 cryptic species of Bemisia tabaci in my country, including 2 invasive species and 13 native species. The cryptic species MED (Mediterranean) and MEAM1 (Middle East-Asia Minor 1) are widely distributed in my country. Native species are only distributed in some areas of our country...

Claims

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Application Information

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IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/6888
Inventor 吕志创冀顺霞王晓迪申晓娜梁林刘万学万方浩
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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