Check patentability & draft patents in minutes with Patsnap Eureka AI!

Insect-resistant gene and use thereof

An insect-resistant gene and gene technology, applied in the direction of genetic engineering, plant gene improvement, application, etc., can solve the problems such as the discount of the biological control effect of brown planthopper, slow insecticidal speed, etc., and achieve the effect of eliminating adverse consequences.

Active Publication Date: 2021-02-23
CHINA JILIANG UNIV
View PDF2 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the pathogenicity of Metarhizium anisopliae to BPH is affected by many factors, including environmental factors, the defense mechanism of the host insect itself, etc., and the insecticidal speed is relatively slow. Such a time-lag effect greatly reduces the biological control effect of BPH. become an important limiting factor for its large-scale application promotion

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Insect-resistant gene and use thereof
  • Insect-resistant gene and use thereof
  • Insect-resistant gene and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] The tested N. lugens population was established in our laboratory. It was raised with TN1 rice seedlings in an artificial climate chamber (temperature 24±1°C, photoperiod 16L:8D, relative humidity 70%±5%) to maintain the population. The wild strain of Metarhizium anisopliae was preserved at 4°C on a PDA (potato dextrose agar medium) slant, and cultured and passaged at 25°C.

[0031] The RNA of N. lugens was extracted and cDNA was synthesized. Specifically, 10 fifth instar nymphs of N. lugens were taken in a mortar and ground evenly with liquid nitrogen. The total RNA of N. lugens was extracted by Trizol method, and the quality and concentration of RNA were detected by micro-volume ultraviolet spectrophotometer (NanoDrop ND-2000, USA) and agarose gel electrophoresis. Using the qualified RNA as a template, use PrimeScript TM 1st Strand cDNA Synthesis Kit (TaKaRa, Japan) kit, synthesized cDNA according to the instructions and stored at -20°C for future use.

[0032] Tak...

Embodiment 2

[0037] The conidia suspension of the wild strain of Metarhizium anisopliae (hereinafter referred to as the wild strain) was used to inoculate adults of N. lugens by spraying method, and the expression of Nlserpin1 of N. lugens was detected after induction at different times. Specifically, the conidia of the wild strain were made into 5×10 0.02% Tween-80 aqueous solution 7 Individuals / mL suspension, inoculate adults of brown planthopper by spray method, the inoculation volume is 1mL, inoculate 3 batches, 100 heads in each batch. Inoculate the same volume of 0.02% Tween-80 aqueous solution as a control. Collect worms at 0h (not inoculated), 6, 12, 24 and 48h after inoculation, disinfect the worms with 75% alcohol surface 3 times, 3min each time, wash 5 times with sterile distilled water, and dry the worms Body spare. According to the cDNA sequence of Nlserpin1 gene, a pair of quantitative PCR primers were designed,

[0038] qS1F: 5'-ACCATGATGAGACAGAAGGGAA-3'

[0039] qS1R: 5...

Embodiment 3

[0044] The Nlserpin1 gene fragment digested by NcoI / BamHI was cloned into the fungal expression vector pAN52-1N, so that it was located between the promoter PgpdA and the terminator TtrpC of the glyceraldehyde triphosphate dehydrogenase gene of Aspergillus nidulans to form pAN52-Nlserpin1. The pET29b-Bar plasmid was digested with XbaI, and the excised glufosinate (PPT) resistance gene bar expression element PgpdA-bar-TtrpC was introduced into pAN52-Nlserpin1, which was also digested with XbaI and dephosphorylated, and obtained by screening The clones with the same orientation of the two expression cassettes of Nlserpin1 and bar genes successfully constructed the binary plasmid pAN52-Nlserpin1-Bar containing the target gene and the resistance marker gene. After the plasmid was linearized by HindIII, it was introduced into the wild strain of Metarhizium anisopliae by PEG-mediated protoplast transformation.

[0045] Eight well-growing transformants were randomly selected on the s...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to the field of biological prevention and treatment, in particular to an insect-resistant gene and a use thereof. The insect-resistant gene is an Nlserpin1 gene, and the gene canremarkably improve the toxicity of metarhizium anisopliae to rice pests. The nilaparvata lugens stal immune negative regulation factor Nlserpin1 gene is used as a research object, the clone identification and biological information analysis are carried out on the Nlserpin1 gene, and therefore a new gene resource is provided for the genetic improvement of insecticidal fungi.

Description

technical field [0001] The invention relates to the field of biological control, in particular to an insect-resistant gene and its application. Background technique [0002] Rice is one of the most important food crops in China, and plays an important role in my country's agricultural production and national economy and people's livelihood, among which pest control is the key link to ensure the safe production of rice. Brown planthopper (Nilaparvata lugens ) is one of the most important rice pests in my country. It not only directly sucks rice juice, lays eggs and destroys rice tissues, causing dryness and lodging, but also spreads rice virus diseases, causing significant losses in rice yields. [0003] At present, the development of resistant rice varieties and the use of chemical insecticides are two important means of controlling BPH. Among them, the use of resistant varieties often results in shortened service life, long cycle and high cost due to the rapid and highly...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N15/12C12N15/80C12N1/15A01N63/30A01P7/04C12R1/645
CPCC07K14/8121C12N15/80A01N63/30Y02A40/146
Inventor 王正亮俞晓平付贤树
Owner CHINA JILIANG UNIV
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com