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Target gene for controlling epilachna vigintioctopunctata and application of target gene for controlling epilachna vigintioctopunctata

A twenty-eight-spot ladybug, target gene technology, applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve the problems of no insecticidal activity, no research on the gene function of the twenty-eight-spot ladybug, and achieve Good effectiveness and sensitivity, environmental friendliness, and easy operation

Active Publication Date: 2019-12-27
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since the use of RNAi technology can specifically inhibit the expression of genes, this technology has been widely used in targeted interference with pest genes to achieve the purpose of pest control. However, there is no research on the gene function of the solanum beetle at home and abroad. Reported target genes without insecticidal activity

Method used

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  • Target gene for controlling epilachna vigintioctopunctata and application of target gene for controlling epilachna vigintioctopunctata
  • Target gene for controlling epilachna vigintioctopunctata and application of target gene for controlling epilachna vigintioctopunctata
  • Target gene for controlling epilachna vigintioctopunctata and application of target gene for controlling epilachna vigintioctopunctata

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] The data processing method of following embodiment: For the result analysis that two types of dsRNA carry out bioassay to ladybug Solanum 28, use Excel 2010 to carry out statistics on the survival rate of ladybug 28, utilize SPSS 19.0 software Cox regression analysis was used to draw the graph, and the difference analysis between different concentrations was analyzed by single factor analysis. For the analysis of changes in the expression of target genes after RNA interference, qPCR data was used in 2 -△△Ctmethod (Ct represents the number of cycles) for calculation. Data analysis was performed using SPSS 19.0 software using one-way analysis of variance. Example 1 Obtaining of dsRNA synthesized by the Lesswright kit of growth and development-related genes

[0032] We constructed the transcriptome library based on the genome of the ladybug Solanum solani, and then based on the constructed transcriptome library, researched and screened the genes related to the growth and...

Embodiment 2

[0050] Example 2 The inhibitory effect of dsRNA on ladybug Solanum solani

[0051] 1. Application of the dsRNA synthesized by the kit in inhibiting the growth and development of the ladybug Solanum solani

[0052] Feeding group of ladybug dsLesswright: 10 1st instar larvae of ladybird solanum were placed in a petri dish with filter paper and humidified cotton balls. Soak circular eggplant leaves with a diameter of 12 mm in dsLesswright solutions with a concentration of 100 ng / μL, 200 ng / μL and 500 ng / μL for 1 min, air-dry them at room temperature for 1 h, and then feed the larvae. Two days after the leaf discs, the larvae were fed with normal eggplant leaves.

[0053] Ladybug dsGFP feeding group: 10 1st instar larvae of Ladybug solani were placed in a petri dish with filter paper and humidified cotton balls. Soak circular eggplant leaves with a diameter of 12 mm in the dsGFP solution synthesized by the kit for 1 min, air-dry them for 1 h, then feed the larvae, replace the le...

Embodiment 3

[0062] Example 3 dsLesswright inhibits the expression of the Lesswright gene in ladybird Solanum solani 1. Experimental method

[0063] On the 2nd and 4th day after starting to feed on dsRNA, the 1st instar larvae of Ladybug solani treated with 250ng / μL dsLesswright and dsGFP were collected, and 3 biological replicates were collected for each treatment. The RNA of the lady beetle was extracted and collected, then reverse-transcribed into cDNA, and diluted 10 times as a template for qPCR. Relative quantitative qPCR analysis was performed with P5 and P6 as primers. The qPCR system is (15 μL) containing 5.25 μL of ddH 2 O, 7.5 μL of 2×SYBR Green MasterMix (BIO-RAD Inc, Hercules, CA), 4 μM primers and 1.0 μL of cDNA first-strand template. The reaction instrument for qPCR was Bio-Rad C1000 Real-Time PCR system (BIO-RAD, USA). The reaction conditions were 95°C for 5 min; 95°C for 10 s, 60°C for 30 s, 39 cycles, 3 technical replicates for each sample, and the reaction was performe...

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Abstract

The invention discloses a target gene for controlling epilachna vigintioctopunctata and application of the target gene for controlling epilachna vigintioctopunctata. According to the target gene for controlling epilachna vigintioctopunctata and application of the target gene for controlling epilachna vigintioctopunctata, a highly lethal gene Lesswright of epilachna vigintioctopunctata is screenedout, moreover, dsRNA which efficiently silences Lesswright is designed, Lesswright has a strong lethal effect on epilachna vigintioctopunctata, then a technology capable of efficiently controlling epilachna vigintioctopunctata is developed, that is to say, by directly feeding epilachna vigintioctopunctata with the target gene dsRNA which has high capability of causing deaths of epilachna vigintioctopunctata, the control purpose is achieved. The technology is convenient to operate, good in effectiveness and sensibility and high in insecticidal efficiency, has multiple advantages of being environmentally friendly and the like, and has excellent application prospects.

Description

technical field [0001] The invention belongs to the technical field of pest control. More specifically, it relates to a target gene for preventing and treating ladybug solani and its application. Background technique [0002] Henosepilachnavigintioctopunctata (Fabricius) belongs to the family Coleoptera and is an important agricultural pest with a wide range of host plants. It mainly damages eggplant, potato and tomato and other solanaceous vegetables. Both the larvae and adults feed on the leaves. They like to cluster on the back of the leaves and feed on the lower epidermis and mesophyll. The damaged leaves usually form irregular transparent spots or perforations. In severe cases, the plants will wilt or even die. Ladybug solani has a wide range of distribution in my country, especially in the south of the Yangtze River. In recent years, due to climate warming, trade development, and the expansion of vegetable cultivation area in protected areas, its food is constantly a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12N15/113C07K14/435A01N57/16A01P7/04
CPCA01N57/16C07K14/43563C12N15/113
Inventor 潘慧鹏吕晶郭威杨春晓郭木娟陈诗敏邱宝利刘卓琦
Owner SOUTH CHINA AGRI UNIV
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