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dsRNA (ribonucleic acid) capable of inhibiting FLPs neuropeptide gene and application thereof

A technology for encoding genes and DNA molecules, applied to dsRNA that inhibits FLPs neuropeptide genes and its application fields, can solve problems such as environmental, human and animal safety hazards, and achieve the effects of reducing infection and impact

Active Publication Date: 2012-02-15
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the prevention and control of root-knot nematode is mainly based on the control of chemical drugs. However, due to its harm to the environment and the safety of humans and animals, it has been gradually canceled by the legislative department.

Method used

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  • dsRNA (ribonucleic acid) capable of inhibiting FLPs neuropeptide gene and application thereof
  • dsRNA (ribonucleic acid) capable of inhibiting FLPs neuropeptide gene and application thereof
  • dsRNA (ribonucleic acid) capable of inhibiting FLPs neuropeptide gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Embodiment 1, the acquisition of dsRNA (Mi-flp-18 dsRNA)

[0035] 1. Cultivation and collection of root-knot nematode incognita

[0036] Meloidogyne incognita was isolated from a greenhouse in Shouguang, Shandong, and the hatched second-instar larvae were inoculated into tomato seedlings (Lycopersicon esculentum cv.) grown in sterilized soil for 30 days. After 2 months, collect the tomato root system, wash it clean, place a 100-mesh sieve of appropriate size in a petri dish, place a piece of filter paper on the sieve, place the picked eggs on the filter paper, add deionized water to cover the egg mass, and leave at room temperature (25° C.) for 72 hours, collect the water under the net, and centrifuge at 5000 rpm for 2 minutes. The collected sediment is the second instar larvae of Meloidogyne incognita.

[0037] 2. Double-strand synthesis

[0038] 1), using RNase Kit (Tiangen) to extract the total RNA of nematodes, using M-MLV (Promega) for reverse transcription to ob...

Embodiment 2

[0078] Embodiment 2, the impact of dsRNA (Mi-flp-18dsRNA) on the second instar larvae of Meloidogyne incognita

[0079] 1. Uptake of dsRNA by second instar larvae of M. incognita

[0080] 1. Reagents

[0081] Fluorescein isothiocyanate (FITC) was used as a marker of absorption efficiency in the test;

[0082] Use soaking buffer to promote the absorption of dsRNA by M. incognita.

[0083] Soaking buffer (dissolve FITC, resorcinol, and spermidine in DEPC water, so that the concentration of resorcinol is 2.5%, and the concentration of spermidine is 150 mM).

[0084] Soaking buffer containing Mi-flp-18dsRNA: the dsRNA aqueous solution (Mi-flp-18dsRNA) obtained by Example 1 is mixed with Soaking buffer, the final concentration of Mi-flp-18dsRNA in the mixed solution is 1mg / ml, and Soaking buffer The concentration is diluted to 0.5 times of the original.

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Abstract

The invention discloses dsRNA (ribonucleic acid) capable of inhibiting FLPs neuropeptide gene and application thereof. The dsRNA provided by the invention is double-chain RNA consisting of nucleotide represented by a sequence 1 in a sequence table and nucleotide represented by a sequence 2 in the sequence table. The experiment in the invention shows that dsRNA (ribonucleic acid) is provided by the invention, 2nd instar larvas of southern plant root-knot nematode are soaked, after RNAi, the tendency migration of southern plant root-knot nematode to a host plant is effectively avoided, the infestation of southern plant root-knot nematode on the host plant is reduced, and the propagation of nematode is also inhibited; and from the point of view of biological safety, dsRNA can not generate functional protein, has the characteristic of specifity, and is capable of reducing the influence on non-target organisms.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to dsRNA for inhibiting FLPs neuropeptide gene and application thereof. Background technique [0002] Root-knot nematode is one of the important pathogenic organisms in the world. It has brought serious losses to the world's agricultural production, and the agricultural economic losses caused by it are as high as 157 billion in the world every year. There are more than 90 kinds of root-knot nematodes that have been reported in the world at present, and southern root-knot nematode (Meloidogyne incognita), Java root-knot nematode (M.javanica), peanut root-knot nematode (M.arenaria) and northern root-knot nematode (M. .hapla) accounted for 95% of the whole root-knot nematodes. Meloidogyne incognita causes a series of problems in facility production and reduces the yield of a large number of commercial crops. In northern my country, root-knot nematodes are the main species, and root-knot ...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/63C12N5/10C12N1/21A01N57/16A01N63/00A01P5/00C12R1/19
Inventor 左元梅董林林李晓林
Owner CHINA AGRI UNIV
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