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Use of nematode chitinase genes to control plant parasitic nematodes

A technology of chitinase and transgenic plants, applied in the field of control of soybean cyst nematodes, can solve problems such as yield loss

Inactive Publication Date: 2010-03-24
BASF PLANT SCI GMBH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Actively feeding nematodes thus steal essential nutrients from the plant, resulting in yield loss

Method used

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  • Use of nematode chitinase genes to control plant parasitic nematodes
  • Use of nematode chitinase genes to control plant parasitic nematodes
  • Use of nematode chitinase genes to control plant parasitic nematodes

Examples

Experimental program
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Effect test

Embodiment 1

[0087] Example 1: Cloning of a gene encoding chitinase from soybean cyst nematode

[0088] The chitinase gene for transformation of soybean was generated by de novo synthesis and then cloned into a minimal vector containing the promoter described below. The DNA sequence of the soybean cyst nematode chitinase was obtained from Genbank accession number AF468679.

Embodiment 2

[0089] Example 2: Vector constructs for transformation

[0090] To assess the function of the cloned chitinase-encoding gene, a gene fragment corresponding to the polynucleotide of SEQ ID NO: 1 was cloned downstream of the promoter to generate the expression vectors described in Table 1. Syncytium-preferred promoters include Arabidopsis pAt5g12170 promoter SEQ ID NO: 3 (US provisional application 60 / 899,693 and PCT / EP2008 / 051329), Arabidopsis TPP 6-phosphate trehalose phosphatase promoter SEQ ID NO: 4 (pAt1g35910) (US provisional application 60 / 874,375 and PCT / EP2007 / 063761). A constitutive Super-promoter (U.S. 5,955,646) was also operably linked to the nematode chitinase polynucleotide of SEQ ID NO:1. The plant selectable marker in the vector was the mutant acetohydroxyacid synthase (AHAS) gene from Arabidopsis conferring tolerance to the herbicide ARSENAL (imazapyr, BASF Corporation, Florham Park, NJ). The mutated selectable marker AHAS gene is driven by the Arabidopsis AH...

Embodiment 3

[0093] Embodiment 3: the preparation of transgenic root and nematode bioassay

[0094] Detection of nematode resistance using a proprietary root explant assay. This assay can be found in commonly owned co-pending application USSN 12 / 001,234, incorporated herein by reference and as described below.

[0095] Clean soybean seeds from soybean cultivars were surface sterilized and then germinated 7 days before Agrobacterium inoculation. The cotyledons were excised for transformation. Immediately after excising the explants from the seedlings, the cut ends were dipped into colonies of Agrobacterium rhizogenes containing the different vector constructs described above. The explants were co-cultured on 1% agar in petri dishes for 6 days. After transformation and co-cultivation, soybean explants are transferred to root induction medium with selection agent.

[0096] Two to three weeks after root induction, the elongated roots were removed and the root explants were transferred to a...

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Abstract

The invention provides transgenic plants with increased nematode resistance which comprise a polynucleotide that encodes a nematode chitinase, seeds of such transgenic plants, expression vectors comprising polynucleotides encoding nematode chitinases, and methods for conferring nematode resistance to crop plants.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of priority to US Provisional Application Serial No. 60 / 894,987, filed March 15,2007. field of invention [0003] The present invention relates to the control of nematodes, particularly soybean cyst nematodes. Disclosed herein are methods of producing transgenic plants with increased nematode resistance, expression vectors comprising polynucleotides encoding functional proteins, and transgenic plants and seeds produced therefrom. Background of the invention [0004] Nematodes are tiny worm-like animals that feed on the roots, leaves and stems of more than 2,000 species of vegetables, fruits and ornamentals, causing an estimated $100 billion in crop losses worldwide. One common nematode is root-knot nematode (RKN), which feeds on roots causing characteristic galls. Other root-feeding nematodes tend to be more host-specific cyst- and root spot-nematode classes. [0005] Nematodes ar...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H5/10A01H5/00
CPCC12Y302/01014C12N15/8285C12N9/24C12N9/2442Y02A40/146
Inventor S·希尔S·莫蒂卡
Owner BASF PLANT SCI GMBH
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