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
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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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