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Cotton cells, plants and seeds genetically engineered to express insecticidal and fungicidal chitin binding proteins (lectins)

A lectin and insecticidal technology, applied in genetic engineering, plant genetic improvement, biocide, etc., can solve the problems of not revealing cotton pests, not providing, etc.

Inactive Publication Date: 2001-07-11
MYCOGEN CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although their data are instructive for some cereal pests, Cavalier et al. do not provide an example of activity against a severe cotton pest
Thus, the work of Cavalier et al., although suggestive, does not reveal that one can control serious cotton pests with a single lectin in purified form.

Method used

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  • Cotton cells, plants and seeds genetically engineered to express insecticidal and fungicidal chitin binding proteins (lectins)
  • Cotton cells, plants and seeds genetically engineered to express insecticidal and fungicidal chitin binding proteins (lectins)
  • Cotton cells, plants and seeds genetically engineered to express insecticidal and fungicidal chitin binding proteins (lectins)

Examples

Experimental program
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Embodiment 1

[0035] The following examples are illustrative of certain embodiments of the inventive subject matter. These examples are illustrative and are not to be construed as limiting the invention in any way. Example 1: Gene

[0036] This study evaluated three different chimeric lectin genes (barley, hevein and nettle). Each chimeric gene contains the cDNA for a specific lectin driven by a promoter active in cotton. For convenience, the CaMV 35S promoter is used, but any promoter proven to be active in cotton, such as the Agrobacterium tumefaciens T-DNA promoter, the Agrobacterium rhizogenes T-DNA promoter or the cotton chlorophyll A / B binding Protein gene promoters (Anderson et al., 1993) are useful. This list is exemplary only and not intended to be inclusive of all promoters. Other useful promoters can be used by those skilled in the art to express barley, hevein and nettle agglutinin in appropriate cotton cells, plants and seeds to control problematic cotton pests such as inse...

Embodiment 2

[0041] Embodiment 2: cotton regeneration

[0042] Methods for establishing and maintaining suspension cultures of cotton embryos are similar to those described by Rangan et al. (U.S. Patent No. 5,244,802, incorporated herein by reference) with further modifications from Rajasekaran et al., 1996 (incorporated herein by reference). For convenience, cotton strain B1654 is used, many other upland or Pima cotton varieties are equally useful, and those skilled in the art can make different variety choices according to the needs of their research projects.

[0043]Treat the seeds with 70% ethanol for 3 minutes, and then treat the seeds with 20% CLOROX solution (1% available chlorine) containing 0.01% surfactant TWEEN-20 for 20 minutes to carry out surface sterilization. (TC agar, Hazleton Biologics, Lenexa, KS) White' The seedlings were grown on s medium (Singh and Krikorian, 1981). First, the embryogenic callus culture was established from the seedling explants according to the me...

Embodiment 3

[0045] Free floating cells and large clumps (≥840 μm) were regularly removed weekly to enrich for small volume, isodiametric, cytoplasmically dense and highly embryonized suspension cultures. Two days before use, these cultures were subcultured in 250 ml Erlenmeyer flasks containing 40 ml maintenance medium. The cell suspension cultures used in our experiments were rapidly growing embryogenic cells, doubling their net weight within 4-6 days (entering logarithmic growth phase after two days of subculture). All cell suspension cultures used for biolistic transformation experiments were cumulatively aged 3-4 months. Example 3: Biolistic transformation of embryogenic cotton cultures

[0046] Three plasmids (barley agglutinin coding region in pGA643, hevein coding region in pGA643, nettle agglutinin coding region in pGA643) were coated with 1.0 μM gold microparticles, followed by a modified helium A driven biolistic device (PDS, 1000 / He; BioRad) was injected into embryogenic cott...

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Abstract

Chimeric genes encoding lectins exhibiting pesticidal activity (for example, insecticidal and / or fungicidal activity) are disclosed which can be used to transform cotton to yield cotton cells, plants, and seeds in which the chimeric genes are expressed. Such transformed cotton cells are pesticidal when ingested by cotton pests.

Description

field of invention [0001] The present invention relates to chimeric genes expressed in cotton cells, plants and seeds that encode insecticides and fungicides substantially possessing the insect and fungal toxicity of barley, nettle and hevein lectins. Background of the invention [0002] Chitin-binding proteins (lectins) are present in many plant species, including monocots and dicots, although these plants do not contain chitin. They are thought to be related to defense, and many of them exhibit insecticidal and / or antifungal activity (Murdock et al., 1990; Lerner, D.R. and Raikhel, N.V., 1992). Lectins exhibit specific carbohydrate-binding properties. Lectins in plants are speculated to be defense-related proteins that exert their action by binding to N-acetylglucosamine of susceptible pest classes (Schroeder, M.R. and Raikhel, N.V. 1992). [0003] Barley, nettle and hevein lectins in purified form exhibit insecticidal and fungicidal activity agains...

Claims

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Application Information

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IPC IPC(8): C07K14/42C12N15/29C12N15/82
CPCA01N65/00C12N15/8286C12N15/8282C07K14/42C12N15/8285A01N65/08A01N65/44Y02A40/146
Inventor R·L·耶诺夫斯基M·范恩T·S·兰根D·M·安德森
Owner MYCOGEN CORP
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