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Rice blast resistant gene RMg1, RMg2 or RMg3, and its application

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

Active Publication Date: 2013-10-09
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, only 14 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 resistant gene RMg1, RMg2 or RMg3, and its application
  • Rice blast resistant gene RMg1, RMg2 or RMg3, and its application
  • Rice blast resistant gene RMg1, RMg2 or RMg3, and its application

Examples

Experimental program
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Effect test

Embodiment 1

[0025] Embodiment one: Cloning and Identification of Rice Blast Resistance Gene RMg1 (Os02g27540-GM)

[0026] 1. Determination of the candidate site RMg1 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;

[0027] 2. Isolation and cloning of rice blast resistance gene RMg1: 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:10; the reverse primer sequence is shown in SEQ ID NO:11.

[0028] 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 ...

Embodiment 2

[0035] Embodiment two: Cloning and Identification of Rice Blast Resistance Gene RMg2 (Os05g23990-Tetep)

[0036] 1. Determination of the candidate site RMg2 for resistance to rice blast: (1) It exists in the form of a single gene, and there is one copy in each of Nipponbare and 93-11; (2) It has a higher LRR region, especially the xxLxLxx region Ka / Ks value;

[0037] 2. Isolation and cloning of rice blast resistance gene RMg2: 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 sequences are listed in the sequence table, the forward primer sequence is shown in SEQ ID NO: 12; the reverse primer sequence is shown in SEQ ID NO: 13.

[0038] 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°...

Embodiment 3

[0045] Embodiment three: Cloning and Identification of Rice Blast Resistance Gene RMg3 (Os08g01580-Q2436)

[0046] 1. Features of the rice blast resistance candidate locus RMg3: (1) It exists as a single gene, with one copy in each of Nipponbare and 93-11; (2) It has a high Ka in its LRR region, especially in the xxLxLxx region / Ks value;

[0047] 2. Isolation and cloning of rice blast resistance gene RMg3: 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.

[0048] 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, denat...

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Abstract

The invention which belongs to the technical field of genetic engineering concretely relates to a rice blast resistant gene RMg1, RMg2 or RMg3, and its application. The invention discloses nucleotide sequences of the new rice blast resistant genes RMg1, RMg2 and RMg3, and amino acid polypeptide sequences coded by the nucleotide sequences. The three genes belong to an NBS-LRR (nucleotide-binding site and leucine-rich-repeat) type disease resistant gene family. The three rice blast resistant genes of the invention are cloned from a rice line highly resisting rice blast pathogens, and are converted to lines which can suffer from the rice blast pathogens, and the disease resistances of the lines are assessed through using a rice blast pathogen invasion method, so a case that the three genes have resistances to the rice blast is finally determined.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to rice blast resistance gene RMg1, RMg2 or RMg3 and application thereof. Background technique [0002] Plant disease resistance genes are the result of long-term interactions between plants and pathogenic bacteria, and play a very important role in the evolution of plants. Since the first plant resistance gene Hm1 was isolated from maize by Johal and Briggs in 1992 (Tanksley et al. 2007; Dong Yuchen, 2001), so far, more than 50 plant resistance genes have been isolated from different plants It was cloned and isolated, and the main type of disease resistance gene was the disease resistance gene of NBS-LRR structure type (Nucleotide-binding site and leucine-rich-repeat, that is, nucleotide binding site and leucine-rich repeat protein); In addition, a small number of disease resistance genes are mainly eLRR-TM-pkinase, eLRR-TM, STK and other types. These dise...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12N15/113C12N15/63C07K14/415A01H5/00A01N47/44A01P3/00
Inventor 田大成王娇杨四海徐婷
Owner NANJING UNIV
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