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Aphid high lethal gene and application thereof in aphid control

An aphid and encoding gene technology, applied in the application, genetic engineering, plant genetic improvement and other directions, can solve the problems of insufficiently high mortality rate of aphid RNA preparations, discounted bioassay effects, difficult dsRNA activity, etc., to reduce the survival rate of melon aphid, The effect of protecting agricultural production and increasing the fatality rate

Pending Publication Date: 2022-05-13
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in previous indoor bioassay studies, the mortality rate caused by gene interference was about 40-60%, and a few genes such as wheat aphid ecdysone receptor gene EcR and supercoiled protein gene USP, soybean aphid immune-related genes hem and energy metabolism The RNA i of the gene ATP, after several days of continuous action, the mortality rate can reach more than 80% (Yan et al., 2016; Yan et al., 2020a; Zheng et al., 2019), however, dsRNA is difficult to maintain in nature and in vivo Long-term activity, such lethal time and lethal rate are far from the insecticidal efficiency of chemical insecticides, and the field test is complicated, which is much lower than the indoor biotest effect, thus restricting RNA as a green insecticide In the actual promotion and application in the field, so far there is no commercial aphid RNA preparation successfully developed and actually used
One of the main reasons why the mortality rate of aphid RNA preparations is not high enough, and it is difficult to match the lethality effect of chemical pesticides over 90%, is that there is no target gene with short action time, low dose and high lethality

Method used

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  • Aphid high lethal gene and application thereof in aphid control
  • Aphid high lethal gene and application thereof in aphid control
  • Aphid high lethal gene and application thereof in aphid control

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Embodiment 1, the acquisition of dsRNA

[0054] 1.1. The source of insects used in this experiment is the melon aphid, which is grown indoors by the Laboratory of High-risk Agriculture and Forestry, China Agricultural University. The indoor breeding conditions are: temperature 27°C; relative humidity kept at about 75%; day: night). The breeding method of the melon aphid is as follows: plant cucumber seedlings, wait for the seedlings to grow 2-3 true leaves, inoculate each leaf with 5 adults of the melon aphid, and reproduce and raise them under the above-mentioned feeding conditions.

[0055] 1.2. Extraction of total RNA of Aphid melon and reverse transcription of cDNA

[0056] Take 5 adult melon aphids, and use the RNAsimple Total RNA Kit (spin column type, kit model DP419) of Tiangen Biochemical Technology (Beijing) Co., Ltd. to extract the total RNA of the sample. Get the RNA of the test sample.

[0057] 1% agarose gel electrophoresis (Leica CM1850) and ultra-micr...

Embodiment 2

[0107] Example 2, the application of dsRNA targeting the synapsin gene in the prevention and treatment of melon aphids

[0108] 2.1. Raw materials for testing

[0109] The nanocarrier SPc is a star-shaped cationic polymer with a structural formula such as Figure 5 Shown, prepare according to the method shown in Example 2 in the Chinese invention patent whose publication number is CN108794710A.

[0110] Alkyl glycoside (APG), a nonionic surfactant that promotes insect body wall penetration, is a commercially available product (Wanhua Guangzhou Supply Chain, product number APG0810), and the dosage is 0.1% of the total volume.

[0111] For testing pest melon aphid is the same as embodiment 1.

[0112] 2.2. Preparation of compound preparation

[0113] 2.2.1, APG solution preparation

[0114] Take 1 μL stock solution and add it into 99 μL sterile water to make 1% APG mother solution for later use. When in use, add 10 μL of 1% APG to every 100 μL of the compound preparation, th...

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Abstract

The invention discloses an aphid high lethal gene and application thereof in aphid prevention and control, and belongs to the field of agricultural pest prevention and control. The invention discloses application of Synapsin or a substance for regulating and controlling the expression of a Synapsin coding gene or a substance for regulating and controlling the activity or content of the Synapsin in regulating and controlling the survival rate of aphids. The Synapsin is a protein with an amino acid sequence shown as a sequence 4 in a sequence table. The invention also provides a preparation for preventing and treating aphids. The preparation contains double-stranded RNA (Ribonucleic Acid) which is Agsyn dsRNA-1 or / and Agsyn dsRNA-2. RNAi application is carried out on aphis gossypii nerve synapse conduction related genes by applying a body wall dropwise addition method, a phenotype with a high fatality rate is obtained on the premise of ensuring interference efficiency, and the fatality rate of dsRNA is improved by applying a nano-carrier.

Description

technical field [0001] The invention belongs to the field of agricultural pest control, and in particular relates to an aphid highly lethal gene and its application in aphid control. Background technique [0002] Aphis gossypii Glover, also known as cotton aphid, is a kind of aphid belonging to Hemiptera Aphidinae (Hemiptera Aphidinae), and is a worldwide omnivorous pest. Melon aphid has a wide range of hosts, including more than 900 species in 116 families, among which it is the most serious damage to melon vegetables, fruits and cotton (Tang Pinghua et al., 2013). The melon aphid has piercing-sucking mouthparts, and it mainly damages host crops by feeding on plant juice, excreting a large amount of honeydew, and spreading viral diseases. For a long time, the control of melon aphid has mainly relied on chemical pesticides, which has led to the continuous development of resistance of the insect, and has become a highly resistant and difficult-to-control aphid species (Carle...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/435C12N15/12C12N15/85C12N5/10C12N15/113A01K67/027
CPCC07K14/43563C12N15/8509C12N15/113A01K67/0275C12N2310/14
Inventor 王丹沈杰王思佳曲雪婷
Owner CHINA AGRI UNIV
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