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Application of phenol sugar acyltransferase gene btpmat1 and its specific dsRNA in the control of whitefly

A technology of bemisia tabaci phenol sugar acyl and transferase, applied in the direction of transferase, DNA/RNA fragments, applications, etc., can solve problems such as food safety threats, environmental pollution, and decreased survival rate of non-target organisms

Active Publication Date: 2021-09-14
INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For a long time, the control of Bemisia tabaci has mainly relied on chemical insecticides. However, the extensive use of chemical insecticides has led to the resistance of Bemisia tabaci to a variety of insecticides, and the long-term use of insecticides has led to non-target A series of problems such as the decline in the survival rate of organisms and environmental pollution pose a serious threat to human food safety.

Method used

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  • Application of phenol sugar acyltransferase gene btpmat1 and its specific dsRNA in the control of whitefly
  • Application of phenol sugar acyltransferase gene btpmat1 and its specific dsRNA in the control of whitefly
  • Application of phenol sugar acyltransferase gene btpmat1 and its specific dsRNA in the control of whitefly

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: Cloning of Bemisia tabaci phenol sugar acyltransferase BtPMaT1

[0027] (1) Discovery of the BtPMaT1 gene in Bemisia tabaci

[0028] Based on the sequencing of the whole genome and transcriptome of Bemisia tabaci, the horizontal gene transfer in Bemisia tabaci was analyzed, and it was found that there was a plant-derived horizontally transferred gene in Bemisia tabaci—the phenol sugar acyltransferase BtPMaT1. The predicted nucleotide sequence of the BtPMaT1 gene was compared with the transcriptome sequence using bioinformatics methods, and the BtPMaT1 gene was corrected.

[0029](2) Cloning the BtPMaT1 gene sequence of whitefly

[0030] Based on the corrected BtPMaT1 gene sequence, the full-length gene-specific primers cBtPMaT1-F and cBtPMaT1-R were designed using Primer Premier 6.0 software. Collect 50 adults of whitefly, use Trizol to extract adult whitefly RNA, and synthesize adult whitefly cDNA with PrimeScript™ II first-strand cDNA kit. The BtPMaT1 ge...

Embodiment 2

[0031] Embodiment 2: the dsRNA preparation of Bemisia tabaci BtPMaT1 gene

[0032] According to the cloned full-length sequence of the BtPMaT1 gene of Bemisia tabaci, specific primers dsBtPMaT1-F and dsBtPMaT1-R containing T7 promoter were designed. Using adult whitefly cDNA as a template, PCR amplification was carried out with the above-mentioned specific primers containing T7 promoter. PCR products were collected by gel recovery and passed through T7 Ribomax TM Express RNAiSystem (Promega, Madison, WI, USA) kit synthesized dsRNA specific for BtPMaT1 gene of whitefly whitefly.

Embodiment 3

[0033] Example 3: Lethal effect of phenolic sugars on whitefly

[0034] Determination of the lethal effect of phenolic sugar plant secondary metabolites on Bemisia tabaci by feeding pouches. The bioassay device consists of a two-way glass tube (inner diameter, 20mm; length, 50mm), a black cotton plug, a black tube sleeve and a double-layer feeding pouch made of parafilm. Add 0.2 ml of feeding solution containing phenolic sugars to the feeding pouch of the bioassay device, seal one end of the glass tube with the feeding pouch, transfer 50 newly emerged whitefly adults to it, and seal the end of the glass tube with a cotton plug. The other end, then cover the glass tube with a black tube sleeve. The bioassay device was placed in a light incubator at 25 °C with a humidity of 80%, and the side of the feeding pouch was placed upwards, facing the light source, and the mortality of adult Bemisia tabaci was measured after 96 hours. The lethal effect of phenol sugar on Bemisia tabaci...

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Abstract

The present invention discloses Bemisia tabaci phenol sugar acyltransferase protein (protein sequence shown in SEQ ID NO.2) and its coding gene BtPMaT1 (The nucleotide sequence is shown in SEQ ID NO.1), and the application of dsRNA, dsRNA can be specifically silenced BtPMaT1 Genes make whitefly more sensitive to phenolic glycoside plant secondary metabolites, meanwhile, transgenic plants expressing dsRNA can specifically silence whitefly BtPMaT1 gene, and causes the death of whiteflies. The invention also discloses BtPMaT1 The gene fragment is used for the synthesis of dsRNA and the construction of transgenic plant expression vectors. The intake of the dsRNA can enhance the lethal effect of phenolic substances on whitefly adults, and feeding on transgenic plants expressing dsBtPMaT1 can lead to the death of whitefly adults. The invention provides a new target for preventing and controlling the whitefly in the field by means of RNA interference and transgene, and has good application prospects.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to the application of a whitefly phenol sugar acyltransferase gene, its specific dsRNA and a transgenic plant expressing the specific dsRNA in the prevention and control of whitefly. Background technique [0002] Bemisia tabaci is a worldwide invasive pest with a wide range of hosts, including more than 600 species of crops and ornamental plants. Bemisia tabaci affects crops worldwide by feeding on phloem juice, secreting honeydew, and spreading various plant viruses. cause serious harm. For a long time, the control of Bemisia tabaci has mainly relied on chemical insecticides. However, the extensive use of chemical insecticides has led to the resistance of Bemisia tabaci to a variety of insecticides. At the same time, long-term use of insecticides has led to non-target A series of problems such as the decline in the survival rate of organisms and environmental pollu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/10C12N15/54C12N15/113C12N15/82A01H5/00A01H6/82
CPCC12N9/1029C12N15/1137C12N15/8218C12N15/8286C12N2310/14
Inventor 张友军夏吉星郭兆将杨泽众吴青君王少丽谢文
Owner INST OF VEGETABLE & FLOWERS CHINESE ACAD OF AGRI SCI
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