Wheat antidisense related gene TaEDR1 and its application

A disease-resistance-related gene, wheat technology, applied in application, genetic engineering, plant genetic improvement and other directions, can solve the problems of wheat disease-resistance gene, wheat disease-resistance-related gene isolation, cloning and less disease resistance response, etc.

Inactive Publication Date: 2006-04-12
HENAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are relatively few researches on the isolation, cloning and disease resistance response of wheat disease resistance genes and wheat disease resistance related genes in the world.

Method used

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  • Wheat antidisense related gene TaEDR1 and its application
  • Wheat antidisense related gene TaEDR1 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0088] Example 1: Cloning process of wheat disease resistance-related gene TaEDR1:

[0089] Wheat TaEDR1 gene was isolated from a new wheat line "99-2439" with high resistance to powdery mildew. Total RNA (ribonucleic acid) was extracted from leaves 22 hours after powdery mildew induction, and mRNA (messenger ribonucleic acid) was further isolated, and mRNA was transcribed into cDNA (complementary deoxyribonucleic acid) by reverse transcription, and the cDNA was used as a template Perform gene cloning.

[0090] A pair of degenerate primers designed according to the Arabidopsis EDR1 gene and its homologs (the sequence is:

[0091] 5′-GCIGTIAAIAAITTI(T / C)TIGA(T / C)CA(G / A)GA-3′ and 5′-GCIAAIGAIGGIC(G / T)IAI(G / A)TTIGG(G / A)TC-3 '), a 627bp long cDNA fragment representing the wheat TaEDR1 gene was obtained by reverse transcription PCR technology. On the basis of this sequence, a 5'-RACE gene-specific primer (sequence: 5'-CGGAGCCGACGCATAATCCTCACTT-3') was designed, and a 2268bp long...

Embodiment 2

[0133] Example 2: Application of the wheat disease resistance related gene TaEDR1 of the present invention in transgenic disease resistance breeding.

[0134] The TaEDR1 gene was excised from the pGEM-T easy vector, or amplified with high-fidelity Taq-DNA polymerase and gene-specific primers, and TaEDR1 was connected to the maize ubiquitin efficient promoter Ubi( The downstream of Ubiquitin-1) was inserted into the pCAMBIA-Bar plant expression vector, thereby constructing the pCAMBIA-Ubi-TaEDR1 expression vector plasmid. The expression vector is transformed into Escherichia coli DH10B bacterial strain for propagation, and the plasmid of the vector is extracted to carry out transgenic disease-resistant breeding. Transform the pCAMBIA-Ubi-TaEDR1 expression vector plasmid into the callus tissue of immature embryos of wheat varieties with high yield and high quality but poor disease resistance through a gene gun (such as Bio-Rad's high-pressure helium gene gun, PDS-1000 / He). Usin...

Embodiment 3

[0135] Example 3: Application of wheat disease resistance-related gene TaEDR1 in designing and synthesizing new disease resistance-related genes.

[0136] Modern DNA synthesis technology has been able to synthesize a full-length gene. Therefore, according to the difference between the TaEDR1 gene and the HvEDR1 gene, the cDNA encoding the 45th amino acid residue of the HvEDR1 gene was synthesized and the 41st amino acid residue of the TaEDR1 gene was synthesized to the end of the cDNA of the gene. Linked together, so that a new gene is synthesized. Then, the correct coding frame (ORF) of the synthetic gene was verified by sequencing. The verified gene can be further constructed into a plant expression vector, and the disease resistance of the synthetic gene can be tested through transgenic technology. The synthetic gene with good disease resistance can be used for the cultivation of new transgenic disease-resistant wheat varieties, and its plant expression The construction of...

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Abstract

A wheat gene TaEDR1 associated with disease resistance is separated from the wheat í‹99-2439íŒ which has high resistance to powdery mildew. The complete length of its cDNA is 3050 bp. The protein coded by said gene is a mitogen protein kinase consisting of 959 amino acids. Said TaEDR1 can be used to improve disease-resistant plant, synthesize new disease-resistant gene and prepare wheat gene chip.

Description

[0001] 1. Technical field: the present invention relates to the technical field of plant genetic engineering, in particular to a kind of wheat disease resistance-related gene TaEDR1 and its application. 2. Background technology: [0002] Plant transgenic research began in the early 1980s. In 1983, Zambryski obtained the world's first transgenic plant. In 1985, Horch created the leaf disk method in Agrobacterium transformation. Since then, Agrobacterium-mediated methods have been established. , Virus-mediated method, PEG-mediated method, electroporation method, microinjection method, pollen tube channeling method, ultrasonic method, gene gun method, etc., the number of successfully transgenic species continues to expand, involving more than 50 species and a total of more than 110 plants Among them, the transgenic plants obtained by the Agrobacterium-mediated method accounted for more than 85% of the total number of transgenic plants, and the transgenic plants obtained by the bio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415C12N9/12C12N15/54C12N15/82
Inventor 牛吉山郭天财张丽娜王映红洪德峰
Owner HENAN AGRICULTURAL UNIVERSITY
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