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MiRNA for regulating lepidoptera pest death, analogue and inhibitor and application thereof

A lepidopteran and analog technology, which is applied in the field of preparation of lepidopteran pest control agents, can solve the problems of overlapping generations, environmental pollution, irrational use of pesticides and the like

Pending Publication Date: 2021-07-13
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Lepidoptera pests are important pests in the production of vegetable crops. Often a variety of pests occur in combination, with many generations in a year and serious overlapping of generations, causing huge economic losses every year.
At present, the main means of controlling lepidopteran pests still rely on the chemical control of insecticides, but due to the unreasonable use of a large number of insecticides, lepidopteran pests have produced almost all the used insecticides. In order to achieve better control effect, we can only continuously increase the use of pesticides, which will cause more serious environmental pollution and form a vicious circle

Method used

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  • MiRNA for regulating lepidoptera pest death, analogue and inhibitor and application thereof
  • MiRNA for regulating lepidoptera pest death, analogue and inhibitor and application thereof
  • MiRNA for regulating lepidoptera pest death, analogue and inhibitor and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] In this example, based on the above-mentioned synthetic pxy-miR-34-5p analog agomiR-34 and inhibitor antagomiR-34, the corresponding pest control effect was verified in beet armyworm, and at the same time, the general nonsense nucleic acid Sequences agomiR-NC and antagomiR-NC were used as controls.

[0035] The above agomiR-34, agomiR-NC, antagomiR-34 and antagomiR-NC were dissolved and diluted to 4 μg / μL with nuclease-free water, respectively.

[0036] Select beet armyworm larvae of the same age, and inject the above-mentioned agomiR-34 / antagomiR-34 into the insects with a microinjector, and the specific standards are as follows:

[0037] (1) Select beet armyworm larvae at the end of the second instar, inject 1.4 μg of agomiR-34 solution into the abdomen of beet armyworm (treatment group), and inject the same amount of agomiR-NC to the control beet armyworm. Raise one head with artificial feed in the feeding cup;

[0038] (2) Select the larvae of Spodoptera exigua in...

Embodiment 2

[0046] In this example, based on the above-mentioned synthesized pxy-miR-34-5p analogue agomiR-34 and inhibitor antagomiR-34, the corresponding pest control effect was verified in cotton bollworm, and the general nonsense nucleic acid sequence agomiR-NC and antagomiR-NC were used as controls.

[0047] The above agomiR-34, agomiR-NC, antagomiR-34 and antagomiR-NC were dissolved and diluted to 4 μg / μL with nuclease-free water, respectively.

[0048] Cotton bollworm larvae of the same age were selected, and the above-mentioned agomiR-34 / antagomiR-34 were injected into the insects with a microinjector, and the specific standards were as follows:

[0049] (1) Select cotton bollworm larvae at the end of the second instar, inject 2.2 μg of agomiR-34 solution into the abdomen of the cotton bollworm (treatment group), and inject the same amount of agomiR-NC to the cotton bollworm in the control group. Raise one head with artificial feed;

[0050] (2) Select cotton bollworm larvae at ...

Embodiment 3

[0058] In this example, based on the above-mentioned synthesized pxy-miR-34-5p analogue agomiR-34 and inhibitor antagomiR-34, the corresponding pest control effect was verified in Plutella xylostella, and the general nonsense nucleic acid sequence agomiR-NC and antagomiR-NC were used as controls.

[0059] The above agomiR-34, agomiR-NC, antagomiR-34 and antagomiR-NC were dissolved and diluted to 4 μg / μL with nuclease-free water, respectively.

[0060] Select Plutella xylostella larvae of the same age, and inject the above-mentioned agomiR-34 / antagomiR-34 into the insects with a microinjector, and the specific standards are as follows:

[0061] (1) Select Plutella xylostella larvae in the middle stage of the third instar, inject 207ng of agomiR-34 solution into the abdomen of Plutella xylostella (treatment group), inject the same amount of agomiR-NC into the control group Plutella xylostella, and after injection, use cabbage leaf feeding;

[0062] (2) Select Plutella xylostel...

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Abstract

The invention belongs to the technical field of agricultural biology, particularly relates to miRNA capable of regulating lepidoptera pest death and delaying growth and development of lepidoptera pests as well as an analogue and an inhibitor of the miRNA, and further discloses application of the miRNA to preparation of medicaments for preventing and treating lepidoptera pests. According to the present invention, the sequence of a lepidoptera insect miRNA gene family is analyzed to obtain the lepidoptera insect plutella xylostella miRNA, i.e., pxy-miR-34-5p, and the mature body sequence of the pxy-miR-34-5p is further labeled and chemically modified so as to synthesize the corresponding miRNA analogue agomiR-34 and the inhibitor anti-agomiR-34 in vitro. A microinjection method is used for researching and finding that the analogue agomiR-34 and the inhibitor agomiR-34 of the pxy-miR-34-5p can delay the growth and development of lepidoptera pests or reduce the survival rate of lepidoptera pests by regulating and controlling the EcR of lepidoptera pests, and theoretical guidance is provided for establishing a new strategy for controlling lepidoptera pests based on the miRNA interference technology.

Description

technical field [0001] The invention belongs to the field of agricultural biotechnology, specifically a miRNA that can regulate the lethality of lepidopteran pests and delay the growth and development of lepidopteran pests, and its analogs and inhibitors, and further discloses its use in the preparation of medicaments for preventing and controlling lepidopteran pests use. Background technique [0002] Lepidoptera pests are important pests in the production of vegetable crops. Many kinds of pests often occur mixedly, with many generations and serious overlapping of generations, which cause huge economic losses every year. At present, the main means of controlling lepidopteran pests still rely on the chemical control of insecticides, but due to the unreasonable use of a large number of insecticides, lepidopteran pests have produced almost all the used insecticides. In order to achieve a better control effect, the use of pesticides can only be continuously increased, which in ...

Claims

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Application Information

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IPC IPC(8): C12N15/113A01N57/16A01P7/04
CPCC12N15/113A01N57/16C12N2310/141
Inventor 朱斌李琳红梁沛高希武
Owner CHINA AGRI UNIV
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