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Nematode-resistant transgenic plants

A technology of transgenic plants and transcription factors, applied in the field of nematode-resistant transgenic plants, can solve the problem of not deregulation of genetically modified plants, etc.

Inactive Publication Date: 2012-05-30
BASF PLANT SCI GMBH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0011] To date, genetically modified plants containing a transgene conferring nematode resistance have not been deregulated in any country

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0079] Embodiment 1: vector construction

[0080]Using the available cDNA sequences of soybean target polynucleotides, PCR was used to isolate DNA fragments that were used to construct the binary vectors described in Table 1 and discussed in Example 2. The PCR product was cloned into TOPO pCR2.1 vector (Invitrogen, Carlsbad, CA) and the insert was confirmed by sequencing. Using this method to isolate polynucleotides GmAnnAt4-like (SEQ ID NO: 49), GmAux28 (SEQ ID NO: 127), GmIsoflavone7OMT-9 (SEQ ID NO: 117), GmAnUGT_47218626 (SEQ ID NO: 109), Gmhsr201- The open reading frames described by MtTINY-like (SEQ ID NO: 105), MtTINY-like (SEQ ID NO: 39), GmLaccase1 (SEQ ID NO: 79) and GmHI1 (SEQ ID NO: 21). Alternatively, available soybean genomic sequences were used to design primers for amplification from the soybean genomic DNA gene sequence to construct the binary vectors described in Table 1 and discussed in Examples 2 and 3. The DNA sequence of the soybean target gene was PCR ...

Embodiment 2

[0085] Example 2: Nematode Bioassays

[0086] Bioassays to assess nematode resistance conferred by polynucleotides described herein were performed using the rooting 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 race3 SCN second stage larvae (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 average cyst number and standard error for 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. Rooted explant cultures transformed w...

Embodiment 3

[0088] Example 3: Root Biomass Determination

[0089] The rooting plant assay system disclosed in co-owned co-pending USSN 12 / 001,234 was also used to assess root growth of uninfected transgenic roots comprising RTP2833, RTP2834, and RTP2839. Multiple transgenic roots and attached cotyledons were subcultured on agar plates for observation. Mark the root tip on the back of the plate as a reference point during subculture. Incubate subcultured roots and cotyledons side by side in the light chamber for up to 6 days. Root weight, root length and number of lateral roots were recorded for each transformation construct. Root parameter measures for each transformation construct were compared to those of the empty vector control tested in parallel to determine whether the tested construct resulted in changes in root weight, root length, root area, and lateral root number. Rooted explant cultures transformed with vectors RTP2833 and RTP2839 showed a general trend of increased root we...

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Abstract

The invention provides nematode-resistant transgenic plants and seed that express polynucleotides encoding AP2 / EREBP transcription factors, harpin-induced proteins, TlNY-like transcription factors, annexins, laccases, isoflavone 7-0-methyltransferases, anthocyanidin 3-glucoside rhamnosyltransferases, hsr201 -like, or AUX / IAA proteins. The invention also provides methods of producing transgenic plants with increased resistance to plant parasitic nematodes and expression vectors for use in such methods.

Description

field of invention [0001] The present invention relates to improving 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, characterized by causing root gall formation at feeding sites, has a relatively broad host range and is therefore parasitic on 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): C12N15/82A01H5/00
CPCC12N15/8261C12N15/8285Y02A40/146
Inventor B·麦凯格A·威格D-S·希尔S·莫蒂卡
Owner BASF PLANT SCI GMBH
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