A rice blast resistance gene rmg41 and its application

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

Active Publication Date: 2017-01-11
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

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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  • A rice blast resistance gene rmg41 and its application
  • A rice blast resistance gene rmg41 and its application
  • A rice blast resistance gene rmg41 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1: Evolutionary analysis of rice blast resistance gene RMg41

[0022] Based on the whole genome sequencing of rice varieties 93-11 and Nipponbare, all NBS-LRR type disease resistance genes in the genome were identified first, and a phylogenetic tree was constructed. On this basis, we selected 20 candidate blast resistance gene loci, one of which is RMg41. This locus has the following characteristics: (1) The gene mostly exists in the form of gene families and gene clusters, with 4 similar copies in the Nipponbare genome and only 1 copy in the 93-11 genome; (2) Its LRR region, especially the xxLxLxx region has a higher Ka / Ks value.

Embodiment 2

[0023] Example 2: Isolation and Cloning of Rice Blast Resistance Gene RMg41 (Os03g63150_GM)

[0024] Using public database sequencing species 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:4; the reverse primer sequence is shown in SEQ ID NO:5.

[0025] 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, denaturation at 95°C for 30 seconds, renaturation at 60°C for 45 seconds, extension at 68°C for 7 minutes, a total of 35 cycles, followed by incubation at 72°C for 10 minutes, and finally constant temperature at 10°C. Subsequently, the PCR products were recovered by tapping rubber.

Embodiment 3

[0026] Example 3: Preparation of the bifunctional base carrier

[0027] 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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Abstract

The present invention belongs to the technical field of gene engineering, and particularly relates to a rice blast resistance gene RMg41 and applications thereof. According to the present invention, the nucleotide sequence and the amino acid polypeptide sequence encoded by the gene of the new rice blast resistance gene RMg41 are disclosed, and the gene belongs to the member of the NBS-LRR type resistance gene family; and the rice blast resistance gene RMg41 is cloned from the rice strain exhibiting high resistance on the rice blast bacteria, the rice blast resistance gene RMg41 is transformed into the rice blast bacterial susceptible variety, and the rice blast bacterial infection method is adopted to evaluate the disease resistance so as to finally determine the resistance of the gene on 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 RMg41 and its application. Background technique [0002] Plant disease resistance genes are the result of long-term interaction between plants and pathogenic bacteria, and play a very important role in the process of plant physiology and evolution. 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 large number of practices have 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). [0003] The traditional breeding of disease-resistant varieties usua...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/113C12N15/84A01H5/00A01N47/44A01P3/00
Inventor 杨四海张彦春张小辉田大成
Owner NANJING UNIV
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