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Plant yellow dwarf resistance-associated protein, coding gene and application thereof

A technology related to resistance and yellow dwarf disease, applied in plant genetic improvement, botany equipment and methods, applications, etc., can solve the problems of difficult exchange separation, positioning, unsuitable separation and cloning, etc., and achieve the effect of improving disease resistance

Inactive Publication Date: 2010-05-12
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, because the genetic relationship between Tripyrum intermedia and wheat is very distant, it is difficult to exchange and segregate between the Chromosome (7X) fragment of Tripyrum intermedia carrying an important gene for resistance to yellow dwarf disease and some homologous chromosomes (7D) of wheat. The genetic distance between Bdv2 and other important yellow dwarf resistance genes and molecular markers on the chromosome (7X) fragment of Tritium intermedium was finely mapped by normal F2 population. disease important gene

Method used

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  • Plant yellow dwarf resistance-associated protein, coding gene and application thereof
  • Plant yellow dwarf resistance-associated protein, coding gene and application thereof
  • Plant yellow dwarf resistance-associated protein, coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Embodiment 1, the discovery of TiNBL1 gene

[0057] 1. Discovery of TiNBL1 specific expression band

[0058] Using cDNA-AFLP technology, one pair of primers was screened out from 256 pairs of primers, which can amplify specific The expression band was absent in 8601(SY), a wheat recurrent parent susceptible to yellow dwarf disease ( figure 1 ; 0, 12, 48, 72 represent BYDV poisonous aphid induction 0 hours, 12 hours, 48 ​​hours, 72 hours). Cloning, sequencing analysis of the specific expression band in the wheat-Thinopyrum intermediary translocation line HW642, found that the specific expression band ( 292bp; disease resistance functional gene TiNBL1 marker fragment) is homologous to the LRR segment of a NBS-LRR protein in rice. The nucleotide sequence of the specific expression band is the 2988th to 3279th nucleotide from the 5' end of Sequence Listing Sequence 2 sequence.

[0059] 2. Obtaining the full-length cDNA sequence of the gene encoding TiNBL1

[0060] On th...

Embodiment 2

[0064] Example 2, Anti-yellow dwarf function analysis of TiNBL1 gene (virus-mediated gene silencing technology)

[0065] 1. Silencing the TiNBL1 gene in HW642

[0066] 1. Prepare the specific expression band of TiNBL1 gene with PacI and NotI restriction sites on both sides respectively (see sequence 3 in the sequence listing). After PacI and NotI digestion, the specific expression band (292bp) of the TiNBL1 gene was inserted between the PacI and NotI restriction sites on the BSMV-γ (γ vector of BMSV virus) by the reverse insertion method, so that the TiNBL1 gene was blocked Driven by the T7 promoter of the γ vector, the recombinant vector BSMV-γ: TiNBL1 was obtained;

[0067] 2. Using the recombinant vector BSMV-γ:TiNBL1 to transform the leaves of the 2-leaf stage seedlings of the wheat-Thinopyrum intermedia translocation line HW642, the specific steps are as follows:

[0068] (1) Inoculate the recombinant vector BSMV-γ:TiNBL1 (or BSMV-γ) on the first and second leaves of th...

Embodiment 3

[0076] Embodiment 3, acquisition of transgenic plants and identification of disease resistance

[0077] 1. Construction of recombinant expression vector pA25-TiNBL1

[0078] pAHC25 contains 2 expression cassettes; the first expression cassette has a maize Ubiquitin promoter, Exon, Intron, GUS, Nos terminators, and both ends of GUS have SmaI and SacI restriction sites; the second expression cassette has a maize Ubiquitin promoter , Exon, Intron, Bar, Nos terminators.

[0079] Using the cDNA of HW642 as a template, PCR amplification was carried out.

[0080] PCR amplification primers:

[0081] TiNBL1-TQSMAI:

[0082] 5'- CCCGGG ATGAAAGCTGCCGAGTCTGCATCA-3'; introduce SmaI restriction site;

[0083] TiNBL1-TQSACI:

[0084] 5'- GAGCTCTC TCAACTCTGCCAATGTTGTGTCGTG-3'; introduce SacI restriction site.

[0085] The PCR reaction program was as follows: first, pre-denaturation at 95°C for 5 minutes; then 35 cycles at 95°C for 30 seconds, 56°C for 1 minute, and 72°C for 1 minu...

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Abstract

The invention discloses a plant yellow dwarf resistance-associated protein, a coding gene and application thereof. The protein provided by the invention is a protein (a) or (b) as follows: (a) the protein consisting of an amino acid sequence shown by a sequence 1 in a sequence table; and (b) the protein which is derived from the sequence 1 by performing substitution and / or deletion and / or addition on the amino acid sequence of the sequence 1 with one or more amino acid residues and is related to resistance to plant yellow dwarf. The invention also discloses the coding gene of the protein, as well as a recombinant expression vector containing the gene. After the coding gene is transferred, the disease resistance of susceptible wheat is greatly improved. The study lays a solid foundation for carrying out study on molecular mechanism of wheat resistance to yellow dwarf and molecular breeding and efficiently cultivating novel varieties of wheat which is resistant to yellow dwarf, high in yield and excellent in quality.

Description

technical field [0001] The invention relates to a plant yellow dwarf resistance-related protein, its coding gene and application. Background technique [0002] Wheat yellow dwarf is a worldwide important viral disease of wheat. Once wheat is infected with yellow dwarf disease, there is no cure, resulting in severe reduction in yield and quality of wheat. Therefore, yellow dwarf disease is also known as "wheat cancer", and it is widely distributed and occurs in all wheat regions in the world. In 1978, due to the outbreak of wheat yellow dwarf disease in the United States, the wheat production was reduced by 60% to 80%. In 1988, the German winter wheat production was reduced by 40% due to the epidemic of yellow dwarf disease. In recent years, Australian wheat has lost about 30 million U.S. dollars every year because of this disease, and yellow dwarf disease has also occurred in New Zealand, Argentina, Turkey, Tunisia and other countries. In 1966, 1970, 1973, 1978, 1980, 1987...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/63C12N15/82C12N5/10C12N1/15C12N1/19C12N1/21C12N15/11A01H5/00
Inventor 张增艳辛志勇赵丹杜丽璞徐惠君李宁黄茜
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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