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Use of trehalase genes to confer nematode resistance to plants

A trehalase, transgenic plant technology, applied in plant products, genetic engineering, botanical equipment and methods, etc., can solve problems such as plant yield loss

Inactive Publication Date: 2009-12-30
BASF PLANT SCI GMBH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The actively feeding nematodes thus steal essential nutrients from the plant, resulting in a loss of plant yield

Method used

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  • Use of trehalase genes to confer nematode resistance to plants
  • Use of trehalase genes to confer nematode resistance to plants
  • Use of trehalase genes to confer nematode resistance to plants

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0101] Example 1: Identification of genes specifically expressed in syncytia

[0102] Microarray analysis of laser-ablated syncytial cells from soybean roots inoculated with second-stage larvae (J2) of the soybean cyst nematode (Heterodera glycines) race3 led to the identification of genes specifically or differentially expressed in syncytia. One such gene (52015943) is called trehalase-like protein. Table 1 summarizes the expression data determined by cDNA microarray analysis of all three cell / tissue samples: syncytia, SCN-infected non-syncytia and untreated control root tissue. Relative levels of gene expression are expressed as normalized signal intensities (± standard deviation) as described above.

[0103] Table 1. Expression of trehalase-like genes

[0104] gene name

Syncytium #1(N)

Syncytium #2(N)

Non-syncytia

control root

52015943 ＊

698±259(4)

525±75(5)

122±38

126±60

[0105] (N) Number ...

Embodiment 2

[0107] Embodiment 2: Cloning of soybean trehalase gene

[0108] The GM59678499 open reading frame was amplified using a standard PCR amplification protocol. Primers used for PCR amplification of trehalase-like sequences are shown in Table 2 and were designed based on the sequence of the open reading frame of GM59678499. The primer sequence (SEQ ID NO: 14) shown in GW59678499F contains an AscI restriction site for easy cloning. The primer sequence shown in SEQ ID NO: 15 contains XhoI for ease of cloning. The primer sequences shown by SEQ ID NO: 14 and SEQ ID NO: 15 (GW59678499F and GW59678499R) were used to amplify the 1674bp open reading frame from the 111th to 1784th bases of SEQ ID NO: 11 (the entire cDNA of GM59678499 sequence).

[0109] Amplified DNA PCR products were confirmed by standard agarose gel electrophoresis and DNA extracted from the gel was TOPO cloned into pCR2.1 using the TOPO TA cloning kit following the manufacturer's instructions (Invitrogen). The clone...

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Abstract

The invention provides transgenic plants that exhibit increased resistance to nematode infection by virtue of overexpression of a gene that encodes trehalase in nematode-induced syncytia. Expression vectors comprising trehalase-encoding polynucleotides and methods of employing such vectors to increase nematode resistance of plants are also provided.

Description

[0001] Cross-references to related applications [0002] This application claims the benefit of priority to US Provisional Application Serial No. 60 / 900,136, filed February 8,2007. field of invention [0003] The present invention relates to the control of nematodes, in particular the present invention relates to the control of soybean cyst nematodes. The present application discloses methods of producing transgenic plants with increased nematode resistance, expression vectors comprising polynucleotides encoding functional proteins, and transgenic plants and seeds produced from the plants. Background of the invention [0004] Nematodes are microscopic worm-like animals that feed on the roots, leaves and stems of more than 2000 types of vegetables, fruits and ornamental plants, causing an estimated $100 billion in crop losses worldwide. A common type of nematode is the root-knot nematode (RKN), whose feeding causes characteristic galls on the roots of a wide range of plant s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H5/00A01H5/10
CPCC12N9/2408C12N15/8285Y02A40/146
Inventor A·威格黄翔S·乔杜里
Owner BASF PLANT SCI GMBH
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