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DsRNA composition for biological control

A composition and nucleotide sequence technology, applied in the direction of microorganism-based methods, microorganism measurement/testing, DNA/RNA fragments, etc., can solve the problems of ecological pollution, slow lethality, unfavorable implementation, etc., and achieve the best economy Effect

Pending Publication Date: 2022-06-07
YUNNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, due to the lack of establishment of insect target gene banks, dsRNA biopesticides mostly target essential genes with high biological conservation to ensure their effectiveness, but they not only target target pests, but also kill other species that express this gene in large quantities , resulting in ecological pollution
In addition, the current dsRNA biopesticides still have a major problem, that is, their lethality is slow or even ineffective, and dsRNA needs to be applied in large quantities many times to ensure high efficiency, which increases the economic cost and application workload of dsRNA pesticides, which is not conducive to the field. Executed in large quantities

Method used

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  • DsRNA composition for biological control
  • DsRNA composition for biological control
  • DsRNA composition for biological control

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0091] 1. Primer design for target genes

[0092] innexin1、innexin2、innexin3、innexin4、DOHH、DHYS、eIF4A、eIF4E、eIF5A、Dip3、PEGS基因的genbank登录号依次为111352120、111354493、111349236、111359390、111352300、022960755.1、1274115148、1274132632、1274128289、111361918、111354038, The gene sequences of SOD49, SOD58 and SOD67 are shown in SEQ ID Nos. 1-3 in sequence.

[0093] The complete sequences of the above innexin1, innexin2, innexin3, innexin4, DOHH, DHYS, eIF4A, eIF4E, eIF5A, Dip3, PGES, SOD49, SOD58, SOD67 genes were used as interference targets, and primer design software Primer Premier 5 was used to design specific primers. The specific primer pairs for the dsRNA sequences are shown in SEQ ID Nos. 4 to SEQ ID No. 31 in sequence.

[0094] 2. Extraction of total RNA and synthesis of cDNA from Spodoptera litura larvae

[0095] Specific steps are as follows:

[0096] 1) Take 10 3-day-old larvae and place them in an EP tube with a small amount of RNAisoTM Plus lysis solution added, fully smash an...

Embodiment 2

[0117] Indoor bioassay of dsRNA composition

[0118] A. Nutrients (amount of 1 portion): 5.0 g of soybean flour, 5.0 g of wheat bran, 2.0 g of yeast extract, 1.0 g of agar powder, and 1.0 g of casein. Accurately weigh the above nutrients and put them in a 200mL crisper.

[0119] B. Mixed powder (amount of 1 part): methylparaben 0.125g, sorbic acid 0.125g, ascorbic acid 0.2g, cholesterol 0.5g, choline chloride 0.0425g, half a grain of Liuhe vitamin pill (crushed). Accurately weigh the above reagents and put them in a 1.5mL centrifuge tube.

[0120] Add 50 mL of distilled water to each part of the nutrients in A, and after high temperature sterilization at 120 ° C for 30 minutes, cool down to 50 ° C, add a part of the mixed powder in B, cool at room temperature to form a solid, and prepare 50 mL of feed.

[0121] The egg pieces of Spodoptera litura that were raised in the same rearing box were taken on the same day, incubated at 27°C and 65% humidity and reared in boxes with 9...

Embodiment 3

[0124] field experiment

[0125] The experiments were carried out in two prevention and control demonstration areas in Songming County and Dehong Mang City, Yunnan Province, respectively, in July and December.

[0126]The corn field was divided into 15 plots of 2 m in length x 2 m in width, and each plot contained about 50 corn plants.

[0127] One month after corn sowing, 100ng / μL, 250ng / μL and 500ng / μL dsRNA compositions were sprayed on the surface of corn leaves at 1L per plot to obtain 25mg / m2 2 dsRNA composition treatment group, 62.5mg / m 2 dsRNA composition treatment group and 125mg / m 2 dsRNA composition treated groups. in ddH 2 O is the blank control, with 125mg / m 2 egfp dsRNA was used as a negative control. After drying for 0.5 hours, Spodoptera frugiperda larvae were randomly inoculated on each maize plant, and each plot included 12 3rd instar larvae, 17 1st instar larvae and one egg mass (about 50 eggs). Investigate the damage of corn after 5 days, and anal...

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Abstract

The invention relates to a dsRNA composition for biological prevention and treatment. The dsRNA composition comprises innexin1 dsRNA, innexin2 dsRNA, innexin3 dsRNA, innexin4 dsRNA, eIF4A dsRNA, eIF4E dsRNA, eIF5AdsRNA, DOHH dsRNA, DHYS dsRNA, Dip3 dsRNA, PEGS dsRNA, SOD49 dsRNA, SOD58 dsRNA and SOD67 dsRNA. The dsRNA composition is characterized in that the dsRNA composition is used for biological prevention and treatment; the dsRNA composition can be used for preventing and treating noctuidae pests.

Description

technical field [0001] The present invention relates to a dsRNA composition for biological control. Background technique [0002] Natural enemies are the most important death factors in all stages of pests. Global food security relies heavily on the control of herbivorous pests. In particular, Spodoptera frugiperda and Spodoptera litura are rampant worldwide, causing severe crop losses. Chemical pesticides are currently the main control methods, but their devastating effects on the environment and other organisms make the use of biological control methods to control agricultural pests inevitable. Natural enemies provide strategies for pest biological control. However, the direct release of natural enemies will not only destroy the ecological balance, but also fail to achieve efficient pest control. Therefore, it is necessary to optimize the prevention and control methods of natural enemies, to avoid excessive release of natural enemies to damage the balance of the ecosys...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/70C12N15/10A01G13/00C12R1/19
CPCC12N15/113C12N15/70C12N15/1003A01G13/00C12N2310/14C12Q2523/301C12Q2523/32Y02A50/30
Inventor 罗开珺蔡成慧蔡秋宸唐红梅孟江慧陈昶旭韩云风张齐周贵芳
Owner YUNNAN UNIV
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