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Rice blast resistance gene pikg, encoded protein and its application

A rice blast and resistance gene technology, applied in the field of rice disease-resistant breeding, can solve environmental pollution, unfavorable sustainable development and other problems, achieve the effect of overcoming chain burden and shortening the period of variety improvement

Active Publication Date: 2021-11-02
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional prevention and control of rice blast mainly relies on fungicides, but long-term use of drugs will cause pathogenic bacteria to develop resistance, and at the same time cause environmental pollution, which is not conducive to sustainable development. Breeding practice shows that the use of rice blast resistance genes to breed broad-spectrum disease resistance New rice variety is economical, efficient and environmentally friendly way to control rice blast

Method used

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  • Rice blast resistance gene pikg, encoded protein and its application
  • Rice blast resistance gene pikg, encoded protein and its application
  • Rice blast resistance gene pikg, encoded protein and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1 Obtaining of rice blast resistance gene Pikg

[0029] The invention utilizes the homologous cloning method to clone the rice blast resistance gene Pikg. In order to excavate and identify new alleles of the rice blast resistance gene Pik, the present invention utilizes the sequences of two constituent genes Pik-1 and Pik-2 of Pik (NCBI accession number: HM048900) as templates to design and amplify the Pik- 1 and Pik-2 exon fragment primers ( figure 1 ,Table 1). Since some studies have shown that Pik-1 can directly bind to rice blast avirulent protein AvrPik-D, and the nucleotide sequence of Pik-2 is relatively conservative, the present invention first selects 3 pairs of primers Ex1 that specifically amplify the exon of Pik-1 -F / R, Ex2-F / R, Ex3-F / R( figure 1 a, 1b), using 79 parts of rice leaf genomic DNA as a template (Table 1), the high-fidelity polymerase KOD enzyme was used to carry out PCR amplification, and the PCR product was detected by electrophoresi...

Embodiment 2

[0037] Example 2 Transplantation of Pikg rice plants and gene function verification

[0038] Since the nucleotide sequences of Pikg-2 and Pike-2 are exactly the same, the present invention connects the DNA fragment of the Pikg-1 coding region with the binary transformation vector pRGV to construct the vector pRGV-Pikg-1, and then transforms the Nipponbare containing Pike-2 Callus, the specific operation is as follows:

[0039] The mature seeds of the Pike-2 transgenic plant Nip-Pike-2 constructed on the background of Nipponbare were used to induce callus on the induction medium, and the EHA105 strain carrying the vector pRGV-Pikg-1 was shaken at 28°C until OD 600 When the value is 0.5-0.6, immerse the rice callus in the bacterial solution for 30 minutes, blot the bacterial solution on the surface of the callus and transfer the callus to the co-cultivation medium for 2-3 days in the dark at 19-22°C . The co-cultured calli were taken out, washed with sterilized deionized water...

Embodiment 3

[0040] Example 3 Resistance characteristics of rice blast resistance gene Pikg

[0041] First of all, in order to compare and clarify whether there are differences in the resistance spectra of the 7 alleles cloned at the Pikg and Pik sites in the present invention, the present invention selected the rice blast strains susceptible to Nipponbare and Lijiang Xintuan black valley Anti-spectrum analysis, these strains originated from Heilongjiang (HLJ), Jiangxi (NC), Hunan (HN) and Hubei (HB) (Table 5). Transgenic plants Nip-Pike (Pike) and Nip-Pikg (Pikg) with Nipponbare as genetic background and monogenic lines IRBLkm-Ts (Pikm), IRBL-CL (Pi1), IRBLks with Lijiang Xintuan Heigu as background -S (Piks), IRBLk-Ka (Pik), IRBLkp-K60 (Pikp) and IRBLkh-K3 (Pikh) were used for resistance identification. Greenhouse spray inoculation identification found that the disease resistance frequency of Pikg was different from the other 7 alleles (Pike, Pik, Pikh, Pikm, Pikp, Piks, Pi1) on the Pik...

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Abstract

The invention discloses a rice blast resistance gene Pikg, encoded protein and application thereof, belonging to the field of rice disease resistance breeding. The rice blast resistance gene Pikg includes Pikg-1 and Pikg-2, and the nucleotide sequence of its coding region and the amino acid sequence of its encoded protein are respectively shown in the sequence table. The gene and protein of the invention can be used to improve the resistance of rice plants to blast fungus.

Description

technical field [0001] The invention belongs to the field of rice disease-resistant breeding, and in particular relates to a rice blast resistance gene Pikg, encoded protein and application thereof. Background technique [0002] Rice (Oryza sativa L.) is an important food crop. More than half of the population in my country takes rice as a staple food. Therefore, maintaining and increasing rice production is of great significance to maintaining food security in our country. However, the yield of rice is affected by various diseases and insect pests. Among them, the fungal disease rice blast, sheath blight and bacterial disease bacterial blight are called the three major diseases of rice due to their serious damage. Among the three diseases, rice blast caused by Magnaporthe oryzae has a high incidence frequency, strong prevalence, and a wide range of impacts. According to statistics, the rice yield loss caused by rice blast is as high as 10%-30% every year. . The traditiona...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/46C12Q1/6895G01N33/68
CPCC07K14/415C12N15/8282C12Q1/6895C12Q2600/13G01N33/68G01N2333/415
Inventor 章志宏孟芬何永刚刘少佳
Owner WUHAN UNIV
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