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RNAi target gene for high efficiency lethality of budworms and application of RNAi target gene

A gene, transgenic plant technology, applied in the fields of biotechnology and agricultural applications, can solve the problems of dsRNA response and target gene silencing efficiency differences, large differences in insect lethal effects, etc.

Active Publication Date: 2019-11-22
CAS CENT FOR EXCELLENCE IN MOLECULAR PLANT SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Different species of insects have different uptake mechanisms for dsRNA, resulting in differences in their response to dsRNA and target gene silencing efficiency. Therefore, the lethal effects of different species of insects are quite different.

Method used

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  • RNAi target gene for high efficiency lethality of budworms and application of RNAi target gene
  • RNAi target gene for high efficiency lethality of budworms and application of RNAi target gene
  • RNAi target gene for high efficiency lethality of budworms and application of RNAi target gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0299] Example 1 Synthesis of Target Gene Sequence and dsRNA

[0300] In order to screen effective aphid target genes based on RNA interference technology, transcriptome sequencing was performed on Myzus persicae and soybean aphid (Aphid glycine) respectively (the sampling and sequencing analysis methods of the two aphids were the same). After extracting total RNA from aphids at different developmental stages, equal amounts of RNA were mixed to form the total RNA of the entire developmental stage of aphids, which was sent to Shenzhen Huada Gene Technology Service Co., Ltd. for transcriptome sequencing using the Illumina Hiseq2000 platform. After removing the adapters from the sequencing results, assemble them with the denove program, and then perform functional annotation on Unigene. In this study, target gene fragments were selected from these functionally annotated Unigenes to amplify and synthesize dsRNA. Through a large number of screening, the present invention amplified...

Embodiment 2

[0311] Example 2 Control effect of target gene dsRNA on aphids

[0312] Inoculate a certain number of green peach aphids or soybean aphids on radish seedlings or soybean seedlings respectively, first record the number of aphids inoculated on each plant, and dissolve the synthesized dsRNA into 2% Tween-80, six The dsRNA concentrations of the target genes are shown in Table 3. Then spray 1ml of dsRNA onto the plants inoculated with aphids, and count the next day as the statistical result of the first day after the dsRNA treatment, then count once every other day, count 3 times in total, and record it as after treatment. Day 1, Day 3 and Day 5 results. 2% Tween-80 and dsGFP were used as controls. The statistical results showed that, compared with the control sprayed with only 2% spit-80, the control effects of 3 target genes of peach aphid and 3 target genes of soybean aphid on the two aphids were all more than 80% ( figure 1 , A, B).

[0313] Table 3. Spray concentration of ...

Embodiment 3

[0316] The population reduction rate statistics of the target gene of embodiment 3 to aphids

[0317] Aphids are parthenogenetic insects, born as 1st instar newborn aphids. The period from the first instar newborn aphid to its ability to give birth is about 5-7 days (affected by the ambient temperature). There is an obvious generational alternation phenomenon in aphids on a plant, that is, different generations and various sizes (different ages) Insects are present at the same time, so when the test plants are inoculated there will be various ages of aphids (such as 2th-4th, and possibly adults), so that after each test treatment, the aphids will soon begin to reproduce and produce offspring. One generation, resulting in an increase in the number of aphids on the tested plants after they were counted before spraying. This is a great interference to the judgment of the control effect of aphid insecticides. Therefore, there is a more strict or relatively accurate calculation me...

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Abstract

The invention relates to an RNAi target gene for high efficiency lethality of budworms and an application of the RNAi target gene, particularly discloses 6 genetic fragments which can cause death of budworm nymphs and / or budworms at the adult stage based on an RNA interference technique, a composition containing dsRNA is sprayed to plants for feeding the budworms or directly sprayed to the surfaces of the budworms, and budworm death can be caused. The method is convenient, quick, accurate and efficient.

Description

technical field [0001] The invention belongs to the field of biotechnology and agricultural application, in particular, the invention relates to an RNAi target gene and its application for efficient lethality to aphids. Background technique [0002] Aphids are an important worldwide pest, belonging to Hemiptera Hemiptera, Aphidoidea Aphidoidea, currently known about more than 4,700 species, small in size, fast in reproduction, and important agricultural and horticultural pests. For the control of aphids, At present, chemical agents are still the main method, but due to its fast propagation speed and strong concealment, its control effect is poor, and it is necessary to use a large amount of pesticides to inhibit its reproduction, which inevitably leads to drug resistance of aphids. [0003] As a tool for gene function research, RNAi has been widely used, especially in animals and plants with imperfect genetic manipulation tools. However, currently in insects, after dsRNA en...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113A01N57/16A01P7/04
CPCC12N15/113A01N57/16C12N2310/14C12N15/63A01N63/60A01N37/46C12N15/8218C12N15/8286C07K14/43504C07K14/43563Y02A40/146A01N63/14C12N15/90C12N15/82
Inventor 苗雪霞李海超关若冰
Owner CAS CENT FOR EXCELLENCE IN MOLECULAR PLANT SCI
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