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dsRNA of wing development related gene vestigial and application thereof in control of citrus fruit fly

A tangerine fruit fly and gene technology, which is applied in the fields of biotechnology and agricultural pest control, can solve problems such as restrictions, and achieve the effects of preventing and controlling invasion and proliferation, ecological security and human health.

Active Publication Date: 2018-04-17
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the control of Bactrocera dorsalis is limited to chemical and physical methods, and the more feasible control method, the sterile male insect technique (SIT), is also limited because it needs to release sterile males year by year

Method used

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  • dsRNA of wing development related gene vestigial and application thereof in control of citrus fruit fly
  • dsRNA of wing development related gene vestigial and application thereof in control of citrus fruit fly
  • dsRNA of wing development related gene vestigial and application thereof in control of citrus fruit fly

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1. Obtaining dsRNA of genes related to wing development

[0053] 1. Samples of Bactrocera dorsalis for testing

[0054]The insect source Bactrocera dorsalis used in this experiment is the indoor population of the Plant Quarantine and Invasion Biology Laboratory of China Agricultural University. The indoor breeding conditions: the temperature is 25°C; the relative humidity is kept at about 70%; D). The breeding method of Bactrocera dorsalis is as follows: Bactrocera dorsalis is reared uniformly in the insect cage, the larval feed (the formula of larval feed is as shown in Table 1) is added to 500ml distilled water, boiled and bottled for subsequent use, and it is frozen after cooling Feed for adults (the feed for adults is obtained by mixing sucrose and soybean peptone in a mass ratio of 1:3) and feed the adults after adding water and moistening. After eating the feed, the larvae mature and pupate in the soil, and about ten days later they emerge as adults, and...

Embodiment 2

[0159] Example 2, the application of the dsRNA of the wing development-related gene vg in the control of Bactrocera dorsalis

[0160] 1. Feeding dsRNA

[0161] The 1st instar healthy and normally developing Bactrocera dorsalis larvae were selected as experimental materials. The experiment was set up with blank control group, dsGFP (irrelevant gene) control group and treatment group. Each group had three replicates, with 40 larvae in each replicate.

[0162] 1. Blank control group (CK)

[0163] Take 20 1.5ml centrifuge tubes, add about 0.15g of crushed Bactrocera dorsalis larva feed and 4 μ lddH to each centrifuge tube 2 O, mix well. Two larvae were placed in each centrifuge tube for feeding. The feeding and growth of larvae were observed daily. After 48 hours, the feed is completely eaten, and then only supplementary feed is sufficient until the larvae grow to the end of the third instar. One of the surviving larvae in the centrifuge tube was picked into the soil to pupat...

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Abstract

The invention discloses dsRNA of wing development related gene vestigial and application thereof in control of citrus fruit fly. A primer is designed for citrus fruit fly development related genes anddsRNA is synthesized first, and application of RNAi is performed on the citrus fruit fly development related genes by adopting a feeding method, so that phenotype of wing developmental malformation is obtained on the premise of ensuring interference efficiency. The dsRNA of wing development related gene vestigial and application thereof in control of citrus fruit fly disclosed by the invention solve the problem that injection implantation of dsRNA is difficul to operate in the field, lay a foundation for future study of directly sprayed RNA preparations to control the citrus fruit fly, and have excellent economic and social benefits for effectively controlling invasion and diffusion of the citrus fruit fly and protecting agricultural production and ecology security and human health in China.

Description

technical field [0001] The invention relates to the fields of biotechnology and agricultural pest control, in particular to the dsRNA of the vestigial gene related to wing development and its application in the control of Bactrocera dorsalis. Background technique [0002] Bactrocera dorsalis Hendel belongs to Diptera, Tephritidae, and Bactrocera, and is an important pest of many tropical and subtropical fruits and vegetables. The expansion situation is grim. Bactrocera dorsalis is an omnivorous pest with a wide range of hosts. It can harm more than 250 kinds of fruits and vegetables in more than 40 families such as citrus, guava, carambola, mango, banana, eggplant, pepper, and melon (Deng Zhisheng, 2001; Shang Hong Sheng, 1997), in 2014, the Ministry of Agriculture of my country listed it as a key agricultural invasion target, and the control strategy of Bactrocera dorsalis has received widespread attention. At present, the control of Bactrocera dorsalis is limited to chemi...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/11A01N57/16A01P7/04
CPCA01N57/16C12N15/113C12N2310/14
Inventor 李志红郭韶堃沈杰
Owner CHINA AGRI UNIV
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