Nematode-resistant transgenic plants

A technology for transgenic plants and transformed plants, which is applied in the field of nematode-resistant transgenic plants, and can solve the problems of not releasing the control of genetically modified plants

Inactive Publication Date: 2011-07-13
BASF PLANT SCI GMBH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0016] To date, genetically modified plants containing transgenes that confer nematode resistance have not been deregulated in any country

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Example 1: Cloning of MtCCP gene and vector construction from Medicago truncatula

[0065]The seeds of Medicago truncatula Jemalong A17 were germinated and cultured in the greenhouse. Genomic DNA was isolated from shoots of these plants, and the MtCCP gene was PCR amplified from this genomic DNA using standard molecular biology techniques. The amplification product was ligated into TOPO entry vector (Invitrogen, Carlsbad, CA).

[0066] The cloned MtCCP gene was sequenced and subcloned into a plant expression vector containing the ubiquitin promoter from parsley (WO 03 / 102198; figure 1 in the p-PcUbi4-2 promoter (SEQ ID NO: 19)). The transformed selectable marker was a mutant form of the acetohydroxyacid synthase (AHAS) selectable gene from Arabidopsis thaliana (Sathasivan et al., Plant Phys. 97:1044-50, 1991), conferring resistance to the herbicide ARSENAL ( Imazapyr, BASF Corporation, Mount Olive, NJ). Expression of AHAS2 was driven by the ubiquitin promoter from p...

Embodiment 2

[0070] Example 2: Nematode Bioassays

[0071] Bioassays to assess nematode resistance conferred by the polynucleotides described herein were performed using the rooted plant assay system disclosed in co-owned co-pending USSN 12 / 001,234. Transgenic roots were generated after transformation with the binary vector described in Example 1. Multiple transgenic root lines were subcultured and inoculated with surface-cleared race 3 SCN second stage juveniles (J2) at a level of approximately 500 J2 / well. Four weeks after nematode inoculation, the number of cysts in each well was counted. For each transformation construct, the number of cysts for each line was calculated to determine the mean number and standard error of the construct. The cyst counts for each transformation construct were compared to the cyst counts for the empty vector control tested in parallel to determine whether the tested construct resulted in a reduction in cyst number. Two independent, biologically replicate...

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Abstract

The invention provides nematode-resistant transgenic plants and seed comprising polynucleotides encoding Medicago truncatula cysteine cluster proteins which comprise no more than four cysteine residues in the respective mature peptides. The invention also provides methods of producing transgenic plants with increased resistance to soybean cyst nematode and expression vectors for use in such methods.

Description

field of invention [0001] The present invention relates to enhancing agricultural productivity through the use of nematode resistant transgenic plants and seeds, and methods of making such plants and seeds. Background of the invention [0002] Nematodes are tiny nematodes that feed on more than 2,000 species of row crops, vegetables, fruits, and ornamentals, causing an estimated $100 billion in crop losses worldwide. A variety of parasitic nematode species infect crop plants, including root-knot nematode (RKN), cyst-forming nematode and lesion-forming nematode. Root-knot nematode, which is characterized by causing root gall formation at feeding sites, has a relatively wide host range and thus parasitizes a variety of crop species. Cyst and lesion forming nematodes have a more limited host range but still cause considerable losses in susceptible crops. [0003] The parasitic nematode is currently found throughout the United States, with the highest concentrations occurring ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/82
CPCC07K14/415C12N15/8285Y02A40/146
Inventor D·P·罗哈S·希尔
Owner BASF PLANT SCI GMBH
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