Rice blast resistance gene RMg4 or RMg5 or RMg6 and application thereof

A rice blast resistance gene and rice technology, applied in the field of genetic engineering, can solve the problems of time-consuming and complicated breeding process, and achieve the effects of broadening the resistance spectrum, shortening the breeding cycle, and enhancing resistance

Inactive Publication Date: 2013-04-24
NANJING UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, only 11 blast resistance genes have been cloned so far
Although the genes that have been located can be utilized by molecular marker-assisted selection, the breeding process is cumbersome and time-consuming. After the new variety is bred, it may be susceptible to the newly generated mutant strain, cloned and directly transformed into a resistant strain. Disease gene may be the most direct and effective way to breed disease-resistant varieties

Method used

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  • Rice blast resistance gene RMg4 or RMg5 or RMg6 and application thereof
  • Rice blast resistance gene RMg4 or RMg5 or RMg6 and application thereof
  • Rice blast resistance gene RMg4 or RMg5 or RMg6 and application thereof

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Effect test

Embodiment 1

[0029] Embodiment one: Cloning and Identification of Rice Blast Resistance Gene RMg4 (Os05g40150-Q2436)

[0030] 1. Determination of the candidate site RMg4 for rice blast resistance: (1) It mostly exists in the form of gene families and gene clusters, and there are two similar copies in each of Nipponbare and 93-11; (2) In its LRR region, especially Yes xxLxLxx region has a higher Ka / Ks value;

[0031] 2. Isolation and cloning of rice blast resistance gene RMg4: Using public database sequencing varieties Nipponbare (Nipponbare) and 93-11 as reference sequences, design primers (both ends of the primers have enzyme cutting sites AscI). The primer sequence is listed in the sequence table, and the forward primer sequence is shown in SEQ ID NO:10; the reverse primer sequence is shown in SEQ ID NO:11.

[0032] Using disease-resistant rice varieties Tetep, Gumei 2 and Q2436 as templates, long-segment PCR (Long-PCR) was used to amplify candidate gene fragments. The PCR program wa...

Embodiment 2

[0040] Embodiment two: Cloning and Identification of Rice Blast Resistance Gene RMg5 (Os08g07390-Q2436)

[0041] 1. Determination of the blast resistance candidate site RMg5: (1) It exists in the form of a single gene, with one copy in each of Nipponbare and 93-11; (2) It has a higher Ka in its LRR region, especially in the xxLxLxx region / Ks value;

[0042] 2. Isolation and cloning of rice blast resistance gene RMg5: Using public database sequencing varieties Nipponbare (Nipponbare) and 93-11 as reference sequences, design primers (both ends of the primers have enzyme cutting sites AscI). Refer to the sequence listing for the primer sequence, the forward primer sequence is shown in SEQ ID NO: 12; the reverse primer sequence is SEQ ID NO: 13.

[0043] Using disease-resistant rice varieties Tetep, Gumei 2 and Q2436 as templates, long-segment PCR (Long-PCR) was used to amplify candidate gene fragments. The PCR program was as follows: pre-denaturation at 95°C for 5 minutes, d...

Embodiment 3

[0051] Embodiment three: Cloning and Identification of Rice Blast Resistance Gene RMg6 (Os11g35210-Tetep)

[0052] 1. Determination of the rice blast resistance candidate locus RMg6: (1) It exists in the form of a single gene, with one copy in each of Nipponbare and 93-11; (2) It has a higher Ka in its LRR region, especially in the xxLxLxx region / Ks value;

[0053] 2. Isolation and cloning of rice blast resistance gene RMg6: Using public database sequencing varieties Nipponbare (Nipponbare) and 93-11 as reference sequences, design primers (both ends of the primers have enzyme cutting sites AscI). The primer sequence is listed in the sequence table, the forward primer sequence is shown in SEQ ID NO: 14; the reverse primer sequence is shown in SEQ ID NO: 15.

[0054]Using disease-resistant rice varieties Tetep, Gumei 2 and Q2436 as templates, long-segment PCR (Long-PCR) was used to amplify candidate gene fragments. The PCR program was as follows: pre-denaturation at 95°C fo...

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Abstract

The invention belongs to the technical field of genetic engineering and specifically relates to a rice blast resistance gene RMg4 or RMg5 or RMg6 and an application thereof. The invention discloses a nucleotide sequence of the rice blast resistance gene RMg4 or RMg5 or RMg6 and an encoded amino acid polypeptide sequence thereof. The three genes belong to members of an NBS-LRR (nucleotide-binding site and leucine-rich-repeat) type anti-disease gene family. The three blast resistant genes disclosed by the invention are cloned from a rice line expressing high resistance to magnaporthe grisea and converted to a magnaporthe grisea-susceptible variety, a magnaporthe grisea infection method is utilized for assessing the resistance against diseases of the genes, and then the three genes can be finally determined to have the resistance to rice blast.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to rice blast resistance gene RMg4, RMg5 or RMg6 and application thereof. [0002] Background technique [0003] Because plant diseases bring huge losses to agriculture and forestry production every year, reasonable and effective prevention and control of plant diseases is one of the key issues that must be solved to ensure the sustainable development of agriculture and forestry. A lot of practice has proved that the development and utilization of existing plant disease-resistant germplasm resources is one of the most effective methods to prevent and control plant diseases (Tanksley et al. 2007; Dong Yuchen, 2001). The traditional breeding of disease-resistant varieties usually involves crossing high-quality and high-yield varieties with materials with strong disease resistance, and then through continuous backcross breeding, finally new varieties with high ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/113C12N15/63A01H5/00A01N47/44A01P3/00
Inventor 杨四海谭生军田大成吴可菁
Owner NANJING UNIV
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