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Insecticidal proteins from plants and methods for their use

An insect protein, insecticidal technology, applied in the field of molecular biology

Active Publication Date: 2020-12-01
PIONEER HI BRED INT INC +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although they have proven to be very commercially successful, these genetically engineered insect-resistant crop plants confer resistance only against a narrow range of economically important insect pests

Method used

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  • Insecticidal proteins from plants and methods for their use
  • Insecticidal proteins from plants and methods for their use
  • Insecticidal proteins from plants and methods for their use

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0441] EXAMPLE 1 - ISOLATION AND IDENTIFICATION OF INSECTICIDAL PROTEINS ACTIVE AGAINST Lepidoptera Species from the Pteridophyte Pteris

[0442]Targeting of soybean armyworm ((SBL)( Insecticidal activity of soybean spider moth)) and corn earworm ((CEW)(corn earworm)). This insecticidal activity exhibits heat sensitivity and protease sensitivity indicative of protein properties.

[0443] Pteris miltiorrhiza (PS930) and Pteris genus (PS995) had similar activity profiles, anion-exchange separated fractions, and similar SDS-PAGE protein profiles at the crude level to crude samples. Due to the limited amount of material, the samples were pooled into one sample for purification steps.

[0444] Combined plant material of PS930 and PS995 was removed from storage at -80°C and kept at liquid nitrogen temperature with Geno / 2010 ball mill (SPEX Sample Metuchen, NJ) grinds it into a fine powder. By adding extraction buffer (50 mM Tris, pH 8.0, 150 mM KCl, 2.5 mM EDTA, 1.5% polyv...

example 2

[0447] Example 2 - Transcriptome sequencing of Pteris albicans and cloning of IPD113Aa

[0448] The transcriptome of a synonym of Pteris albolineata, Pteris albolineata (ID#PS930), was prepared as follows. For total RNA isolation by using Kit for the isolation of total RNA from frozen tissues. Use from Inc. (San Diego, CA) TruSeq TM mRNA-Seq Kits and Protocols Prepare sequencing libraries from the resulting total RNA. Briefly, mRNA was isolated via attachment to oligo(dT) beads, fragmented to an average size of 180 nt, reverse-transcribed into cDNA by random hexamer priming, end repaired, 3'A tailed, and Indexed TruSeq TM Adapter connection. use TruSeq TM The ligated cDNA fragments were amplified by PCR with primers, and the Agilent Quality and quantity of purified PCR products were checked on a DNA 7500 chip. After qualitative and quantitative assessment, by using dual specificity nuclease (DSN) ( Moscow, Russia) process to normalize the 100 ng transcript ...

example 3-I

[0451] Example 3 - Expression of IPD113Aa on transient leaf tissue and insect bioassays

[0452] In order to confirm the activity of the IPD113Aa polypeptide (SEQ ID NO: 1), the corresponding gene (SEQ ID NO: 127) was cloned into the viral promoter dMMV (Dey et al., (1999) PlantMol. Biol. [Plant Molecular Biology] 40 :771-782) in a transient expression system under control. Agrobacterium strains containing the IPD113Aa expression construct were infiltrated into leaves. The Agroinfiltration method of introducing a suspension of Agrobacterium cells into intact plant cells so that superinfection and subsequent plant-derived transgene expression can be measured or studied is well known in the art (Kapila et al., (1997) Plant Science [Plant Science] 122:101-108). Briefly, single leaves of bush bean (common bean (Phaseolus vulgaris)) were agro-infiltrated with standardized bacterial cell cultures of test and control strains. Leaf discs were excised from each plantlet and treate...

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Abstract

Compositions and methods for controlling pests are provided. The methods involve transforming organisms with a nucleic acid sequence encoding an insecticidal protein. In particular, the nucleic acid sequences are useful for preparing plants and microorganisms that possess insecticidal activity. Thus, transformed bacteria, plants, plant cells, plant tissues and seeds are provided. Compositions areinsecticidal nucleic acids and proteins of bacterial species. The sequences find use in the construction of expression vectors for subsequent transformation into organisms of interest including plants, as probes for the isolation of other homologous (or partially homologous) genes. The pesticidal proteins find use in controlling, inhibiting growth or killing Lepidopteran, Coleopteran, Dipteran, fungal, Hemipteran and nematode pest populations and for producing compositions with insecticidal activity.

Description

[0001] cross reference [0002] This application claims the benefit of U.S. Provisional Application No. 62 / 642642, filed March 14, 2018, which is hereby incorporated by reference in its entirety. [0003] References to Sequence Listings Submitted Electronically [0004] An official copy of this Sequence Listing is submitted electronically via EFS-Web as a Sequence Listing in ASCII format under the filename "RTS21844WOPCT_SequenceListing", created on January 21, 2019, and has a size of 1,539 kilobytes and is at the same time as this specification submit. The Sequence Listing contained in said file in ASCII format is part of this specification and is hereby incorporated by reference in its entirety. technical field [0005] The present disclosure relates to the field of molecular biology. Novel genes encoding pesticidal proteins are provided. These pesticidal proteins and nucleic acid sequences encoding them can be used in the preparation of pesticidal agents and in the prod...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N65/04A01N37/46A01N57/16C07K14/415C12N15/09C12N15/52C12N15/62C12N15/82A01P7/04
CPCA01N65/00C07K14/415A01N37/46C12N15/8286C07K2319/00C07K2319/02Y02A40/146A01N63/50
Inventor 刘璐A·伦A·S·昂E·谢珀斯I·乌兰什基钟晓红
Owner PIONEER HI BRED INT INC
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