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Soybean disease-resistant gene and its coded protein and uses

A disease-resistant gene, soybean technology, applied in application, genetic engineering, plant genetic improvement, etc., can solve the problems of rapid mutation of pathogenic bacteria, limited role of conventional breeding, and lack of antigens.

Inactive Publication Date: 2007-06-27
THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the rapid mutation of pathogenic bacteria and the lack of available antigens, the role of conventional breeding is largely limited

Method used

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  • Soybean disease-resistant gene and its coded protein and uses
  • Soybean disease-resistant gene and its coded protein and uses
  • Soybean disease-resistant gene and its coded protein and uses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment 1, the cloning of GmeR

[0044] plant material:

[0045] For the soybean variety "Kehe" No. 1, when the seedlings were 4 weeks old, the whole plant was sprayed with 2.0mM salicylic acid for 48 hours.

[0046] 1. Primer design for GmeR 5’Race

[0047] According to the muskmelon eR gene sequence published in the gene bank (www.ncbi.nlm.nih.gov), the sequence alignment was carried out on the Internet, and it was found that there was a 465bp DNA in the EST sequence of soybean induced by 2.0mM salicylic acid for 48 hours Fragment, up to 83% homology with the 3' end sequence of the melon eR gene, it is speculated that this sequence may be the 3' end gene sequence of the homologous gene in soybean, and this homologous gene in soybean is named as GmeR, and the The 3'-end PCR primers P1 and P2 of GmeR5'Race, and the 5'-end primers P3 and P4 are provided by the GeneRacer kit (Cat.No.L1502-01) of Invitrogen, and the sequences of the above primers are as follows:

[0...

Embodiment 2

[0118] Embodiment 2, the construction of GmeR plant expression vector pGmeR121

[0119] 1. Construction of intermediate vector pCV121

[0120] The gene GmeR expression cassette constructed in this example includes the following gene expression regulatory elements: 35S promoter at the 5' end, gene GmeR and NOS terminator at the 3' end. First construct the intermediate vector pCV121 without the gene GmeR, the construction process is shown in Figure 5, and the specific process includes the following steps:

[0121] 1. Construction of intermediate vector pUC121

[0122] Plasmid vector pBI 121 (Beijing Baierdi Biotechnology Co., Ltd.) was double-digested with restriction endonucleases Sac I and EcoRI, and the 260 bp T-NOS terminator fragment was recovered, which was double-digested with the same enzymes. The vector pUC19 was ligated to obtain an intermediate vector named pUC121, which was identified by restriction endonuclease Sac I and EcoR I, and the results of enzyme digestion...

Embodiment 3

[0129] Embodiment 3, identification of disease resistance of GmeR transgenic tobacco

[0130] The plant expression vector pGmeR121 constructed in Example 2 was transformed into Agrobacterium tumefaciens LBA4404 by freeze-thaw method. The Agrobacterium tumefaciens LBA4404 transformant integrated with pGmeR121 was then transformed into tobacco NC89 by leaf disc method to obtain 20 GmeR transgenic tobacco strains. Using the isolated leaf inoculation method, with non-transgenic tobacco as a control, the genetically modified tobacco is carried out to identify the resistance of tobacco black shank (Black shank) and bacterial wilt. The specific methods are as follows:

[0131] 1. Identification of tobacco black shank resistance in GmeR transgenic tobacco

[0132] The production method of black shank fungus valley: boil the millet until about half of the grains bloom, remove it, put it into a triangular flask, and sterilize it under high pressure for 1 hour. Inoculate the cultured b...

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Abstract

The invention provides enzymatic disease resistance genes of soybean, coded protein and use, wherein the genes are one of the following nucleic acid sequences, (1) DNA sequence of SEQ ID No.1 in the sequence table, (2) DNA sequence of SEQ ID No.2 in the sequence table, (3) nucleic acid sequence capable of hybridizing with the DNA sequence defined by SEQ ID No.1 in the sequence table under highly stringent conditions. The coded protein is one of the following amino acid residue sequences: (1) SEQ ID No.2 in the sequence table, (2) protein obtained through substitution and / or deletion and / or addition of one or several amino acid residuals of SEQ ID No.2 in the sequence table and capable of improving plant disease resisting property.

Description

technical field [0001] The invention relates to a plant gene and its coded protein and its application, in particular to a soybean-derived disease-resistant gene with aminotransferase function and its coded protein and its application in cultivating plants with improved disease resistance. Background technique [0002] Bacteria and fungi are the main pathogenic bacteria that harm crops. Although effective control methods have been found for some diseases, there are no practical and effective control measures for most diseases that seriously endanger the growth of crops. Breeding disease-resistant varieties is the fundamental measure to prevent and control plant diseases. However, due to the rapid mutation of pathogenic bacteria and the lack of available antigens, the role of conventional breeding is largely limited. With the rapid development of molecular biology, plant pathology and genetic engineering technology, the use of genetic engineering to improve plant disease re...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/63C12N15/82
Inventor 贾士荣刘昱辉孙文丽
Owner THE INST OF BIOTECHNOLOGY OF THE CHINESE ACAD OF AGRI SCI