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Nucleotide sequence of gene for resistance to Cereal cyst nematode, Heterodera avenae and application thereof

A nucleic acid sequence and root nematode technology, applied in the field of plant genetic engineering, can solve the problems of unreported functional verification research

Inactive Publication Date: 2011-11-09
CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, only one cereal cyst nematode resistance gene Cre3 has been isolated from Glycyrrhiza japonica through map-based cloning, but its functional verification and other follow-up studies have not been reported

Method used

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  • Nucleotide sequence of gene for resistance to Cereal cyst nematode, Heterodera avenae and application thereof
  • Nucleotide sequence of gene for resistance to Cereal cyst nematode, Heterodera avenae and application thereof
  • Nucleotide sequence of gene for resistance to Cereal cyst nematode, Heterodera avenae and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0025] 1. CreZE Gene complete coding region cloning and sequencing

[0026] 1.1 CreZE Full-length gene sequence amplification

[0027] Primers were designed according to the spliced ​​gene sequence:

[0028] P1: 5'-GGACTAGTCTATCTGTCTGTAGTGATGGAT-3'

[0029] P2: 5'-AAAGCTGGGATTCCAAGCCCGCGTCATTC-3' is used for the amplification of the full-length gene sequence;

[0030] PCR amplification system:

[0031] E10 genomic DNA 2 μL

[0032] 10×LA PCR Buffer 5 μL

[0033] dNTP (2.5mmol / L each) 4 μL

[0034] Forward / reverse primer (10μmol / L) 3 μL each

[0035] TaKaRa LA TaqDNA (5U / μL) 1 μL

[0036] Mg 2+ (25mmol / L) 3 μL

[0037] Add dH 2 0 to 50 μL.

[0038] Reaction cycle conditions: 94°C for 5 min; 94°C for 1 min, 60°C for 1 min, 72°C for 2.5 min, 35 cycles; 72°C for 7 min; 4°C for heat preservation.

[0039] 1.2 PCR product detection and recovery

[0040] (1) Add 50 μL of PCR product and DNA loading buffer into the sample well of 1.2% agarose gel, and electrophoresis...

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Abstract

The invention, relating to the field of plant genetic engineering, particularly relates to a nucleotide sequence of a gene from an allied specie Aegilops variabilis of wheat for resistance to Cereal cyst nematode, and an application on improving plant resistance to insects based on the gene through the method of biotechnology. The nucleotide sequence of the gene for resistance to Cereal cyst nematode disclosed in the invention is a complete encoding sequence of the gene for resistance to Cereal cyst nematode with the width of 3586bp, comprising 3 exons and 2 introns, and can code 934 amino acids, and is used in improving plant resistance.

Description

technical field [0001] The present invention relates to the field of plant genetic engineering, in particular to a nucleic acid sequence of a gene for resistance to root cyst nematode graminearum from a close relative of wheat, Goatweed vulgaris, and the application of the gene in improving plant resistance to insects through biotechnology methods . technical background [0002] Cereal cyst nematode, Heterodera avenae ,) is an important soil-borne pathogenic nematode that harms cereal crops such as wheat. It is distributed in the main wheat producing areas in North my country, and the damage area is more than 1 million hectares. In severe areas, it can cause a loss of 27% to 45% of wheat production. Therefore, it is imminent to discover the genetic resources of resistance to cereal cyst nematodes and to breed new varieties of resistant wheat. [0003] Gene resources for resistance to cereal cyst nematodes in common wheat are very scarce. Cloning and transferring resistanc...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415A01H5/00
Inventor 余懋群徐德林龙海邓光兵潘志芬
Owner CHENGDU INST OF BIOLOGY CHINESE ACAD OF S
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