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Soybean cyst nematode RNA helicase CGH-1, and coding gene and application thereof

A technology of soybean cyst nematode and gene, applied in the direction of DNA/RNA fragments, application, recombinant DNA technology, etc., can solve the problems of cumbersome backcrossing procedures, complicated resistance identification, and restrictions on the breeding of new varieties resistant to soybean cyst nematodes , to achieve the effect of inhibiting infection and transmission, and strong anti-insect activity

Inactive Publication Date: 2012-03-21
INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the use of highly toxic insecticides such as DDT, carbofuran, and aldicarb can kill nematodes, they will cause serious environmental pollution, so they have been banned or restricted.
The problems encountered in cultivating soybean resistant varieties using traditional breeding programs are cumbersome backcrossing procedures and complicated resistance identification
It takes at least ten years to breed a variety through backcross breeding, which greatly limits the selection of new varieties resistant to soybean cyst nematodes

Method used

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  • Soybean cyst nematode RNA helicase CGH-1, and coding gene and application thereof
  • Soybean cyst nematode RNA helicase CGH-1, and coding gene and application thereof
  • Soybean cyst nematode RNA helicase CGH-1, and coding gene and application thereof

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Embodiment 1

[0041] Embodiment 1, the discovery of soybean cyst nematode SCN cgh-1 protein and its coding gene

[0042] 1. Selection of soybean cyst nematode SCN cgh-1 protein and its coding gene

[0043] In the NCBI database, as of May 2009, there were 108,045 related sequences of soybean cyst nematode, including 81,873 nucleotide sequences, 26,172 EST sequences, 183 mRNA and 252 protein sequences. Alkharouf et al processed 24438 EST sequences and 231 genome sequences of soybean cyst nematode, and compared them with the gene sequences of Caenorhabditis elegans, and found that there were 8334 gene sequences similar between the two, and among them, 1508 Genes with lethal phenotype in C. elegans, these genes with lethal phenotype were grouped according to the degree of sequence similarity, and were divided into 6 groups, among which there were 150 genes with the most similar (E-values ​​between 0 and 1E-100). The inventor compared the genetic relationship between 150 genes and humans, and s...

Embodiment 2

[0076] Example 2, Functional Verification of SCN cgh-1 Protein

[0077] 1. Construction of SCN cgh-1RNAi expression vector

[0078] 1. Extract total RNA from the cyst of soybean cyst nematode (physiological race No. 4), and reverse transcribe it into cDNA.

[0079] 2. Using cDNA as a template, carry out PCR with a primer pair consisting of Primer-F and Primer-R (the target sequence is sequence 2 in the sequence listing from 523-1090 nucleotides at the 5' end, i.e. the SCN cgh-1 gene fragment) Amplify to obtain PCR amplification products.

[0080] Primer-F: 5'-CGC TCT AGA GAG CTC CGT CCG ATG GAA GAT TG-3' (Xba I digestion recognition site and Sac I restriction recognition site are underlined);

[0081] Primer-R: 5'-ACG C GT CGA CGG TAC C GG AAG TCC TGG GAA AGG-3' (Sal I digestion recognition site and Kpn I restriction recognition site are underlined);

[0082] PCR amplification system (50 μl): cDNA 1 μl, 10×buffer 5 μl, dNTP (10 mM) 1 μl, Primer-F 1 μl, Primer-R 1 μl,...

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Abstract

The invention discloses a soybean cyst nematode RNA helicase CGH-1, and a coding gene and application thereof. The protein disclosed by the invention is (a) or (b) as shown below: (a) a protein composed of an amino acid sequence shown in sequence 1 in a sequence list; and (b) a protein derived from sequence 1, which is formed by the substitution and / or deletion and / or addition of one or more amino acid residues in the amino acid sequence shown in sequence 1 and is related to the growth of soybean cyst nematodes. In the invention, the soybean cyst nematode RNA helicase CGH-1 and the coding gene thereof are provided, and an RNAi expression vector is designed on the basis of the coding gene; and the RNAi expression vector can be introduced into a plant, possibly the dsRNA of the gene in the plant, so that the nematodes can suffer from severe reproduction or growth dysfunction after taking in the transgenic plant, thereby inhibiting the infection and spread of the nematodes. By using the method disclosed by the invention, a transgenic plant having very high nematode resistance activity can be obtained, thereby laying a foundation for the further cultivation of new broad-spectrum nematode-resistant soybean varieties.

Description

technical field [0001] The invention relates to a soybean cyst nematode RNA helicase CGH-1 and its coding gene and application. Background technique [0002] Soybean originated in China and is an important source of human protein and edible oil. 31% of the world's edible oil comes from soybean. my country is a big consumer of soybeans, with a total annual consumption of more than 40 million tons, while the total annual domestic soybean output is less than 17 million tons. Therefore, it is necessary to rely on a large number of imports to fill the gap. The main problem of my country's soybean production is the low yield per unit area. The average yield per unit area in 2008 was only 114 kg / mu, which was only 70% of the world's average yield. Compared with the United States and other advanced countries, the gap was even greater. In addition to problems such as varieties and planting, diseases and insect pests are important reasons for the low yield of soybeans in my country....

Claims

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Application Information

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IPC IPC(8): C12N9/00C12N15/52C12N15/63C12N5/10C12N1/15C12N1/19C12N1/21A01H5/00
Inventor 杨崇林张劲松陈受宜李海潮荆玉栋蹇友理李金英王昉李锦锦
Owner INST OF GENETICS & DEVELOPMENTAL BIOLOGY CHINESE ACAD OF SCI
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