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Rice blast resistance gene pair TP22 and application thereof

A rice blast resistance gene, TP22 technology, applied in the field of genetic engineering, can solve problems such as ignoring disease resistance genes, and achieve the effect of expanding the resistance spectrum, enhancing resistance, and improving resistance

Inactive Publication Date: 2019-04-19
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

, and in the Tetep genome, we found that more than 20% of the NBS-LRR genes exist in pairs, which suggests that our traditional thinking on the use of a single disease resistance gene ignores the combination and cooperation of disease resistance genes to exert stronger resistance performance mechanism, which may also be the reason why few lines improved by cloning technology can show broad-spectrum and long-lasting resistance to blast fungus

Method used

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  • Rice blast resistance gene pair TP22 and application thereof
  • Rice blast resistance gene pair TP22 and application thereof
  • Rice blast resistance gene pair TP22 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Identification of Rice Blast Resistance Gene TP22

[0036] We searched for NBS-LRR gene pairs with a "head-to-head" structure in the Tetep genome, and constructed phylogenetic trees for their sequences and their homologous sequences in the Nipponbare, Tetep, and Brachypodium genomes to further confirm the pair relationship. A total of 43 disease-resistant genes were obtained in Tetep, and TP22 (chr11.fgenesh2445 / chr11.fgenesh2446) was one of the pairs, which had the following characteristics: (1) chr11.fgenesh2445 (sensing gene) and chr11.fgenesh2446 ( Auxiliary genes) belong to different branches with faster and slower evolution in the phylogenetic tree. (2) chr11.fgenesh2445 (sensing gene) has higher nucleotide polymorphism (π) and nucleotide difference (Dxy) than chr11.fgenesh2446 (auxiliary gene).

Embodiment 2

[0038] Isolation and Cloning of Rice Blast Resistance Gene TP22 (chr11.fgenesh2445 / chr11.fgenesh2446)

[0039] Using the Tetep genome and public database sequencing species Nipponbare as reference sequences, design cloning primers (both ends of the primers have enzyme cutting sites AscI) and fusion primers. For the primer sequence, refer to the sequence listing, the forward cloning primer sequence of chr11.fgenesh2445 is shown in SEQ ID NO: 7; the reverse cloning primer sequence is shown in SEQ ID NO: 8; the forward cloning primer sequence of chr11.fgenesh2446 is shown in SEQ ID NO Shown in: 9; Reverse cloning primer sequence is shown in SEQ ID NO: 10. The forward fusion primer sequence of chr11.fgenesh2445 is shown in SEQ ID NO:11; the reverse fusion primer sequence is shown in SEQ ID NO:12; the forward fusion primer sequence of chr11.fgenesh2446 is shown in SEQ ID NO:13; The reverse fusion primer sequence is shown in SEQ ID NO:14. Using the disease-resistant rice variety T...

Embodiment 3

[0041] Preparation of bifunctional base vectors

[0042] The rare restriction site AscI was introduced between the multiple cloning sites BamHI and SalI of the bifunctional vector pCAMBIA1300, and the restriction site XbaI was replaced to form the basic vector pCAMBIA1300-AscI with the rare restriction site AscI.

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PUM

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Abstract

The invention discloses a rice blast resistance gene pair TP22 and application thereof, and discloses nucleotide sequences and coded amino acid polypeptide sequences of two genes in the rice blast resistance gene pair TP22. The two genes in the gene pair both belong to members of NBS-LRR type disease-resistant gene family. The rice blast resistance gene pair TP22 and the application thereof have the advantages that the two genes resistant to the rice blast are cloned from rice strains showing high resistance to magnaporthe oryzae, and are converted into strains susceptible to magnaporthe oryzae, and the disease resistance of the two genes is evaluated by using a magnaporthe oryzae infection method, thereby finally determining that the gene pair has resistance to the rice blast.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a rice blast resistance gene pair TP22 and its application. Background technique [0002] Rice is an important food crop in the world, feeding more than 3 billion people in the world, and it is also one of the most important food crops in my country. However, 10-30% of rice production is lost annually due to rice blast (Skamnioti and Gurr 2009; Miah et al. 2012; Imam et al. 2013). As the deadliest disease of rice, its pathogen, Magnaporthe grisea can infect rice in almost all growth and development stages, and has the characteristics of long attack time, more symptoms than wide infection sites. Rice blast can be classified according to when it infects rice and where it infects. Such as seedling blast, leaf blast, leaf pillow blast, festival blast, panicle blast, and grain blast. [0003] Plants in nature, including rice, have evolved a large number of d...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/113C12N1/21C07K14/415A01H5/00A01H6/46C12R1/01
CPCC12N15/8282C07K14/415
Inventor 田大成王观杨四海张小辉
Owner NANJING UNIV
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