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Transcription factor for enhancing broad-spectrum disease resistance of plants and application

A plant, disease resistance technology, applied in the field of molecular biology or botany, can solve the problem of poor understanding of regulatory mechanisms and involved transcription factors

Active Publication Date: 2020-09-15
CAS CENT FOR EXCELLENCE IN MOLECULAR PLANT SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In this field, although studies have shown that the interaction between NLRs and transcription factors directly regulates the expression of defense genes during the ETI response, the specific regulatory mechanism and the involved transcription factors are still poorly understood

Method used

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  • Transcription factor for enhancing broad-spectrum disease resistance of plants and application
  • Transcription factor for enhancing broad-spectrum disease resistance of plants and application
  • Transcription factor for enhancing broad-spectrum disease resistance of plants and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0231] Example 1, Screening and Analysis of PigmR Interacting Protein PIBP1

[0232] The present inventor identified a novel site Pigm with broad-spectrum and persistent blast resistance through map-based cloning. Pigm is a gene cluster encoding multiple NBS-LRR disease resistance genes, and the encoded functional protein PigmR has broad-spectrum disease resistance. In order to explain the broad-spectrum disease resistance mechanism of PigmR, the inventors carried out a yeast screening library, and utilized the pDEST32 / pDEST22 vector system of the Gateway system of Invitrogen Company to incorporate the PigmR-CC domain ( figure 1 A) Constructed into pDEST32 as a bait library (rice cDNA yeast library after inoculation of Magnaporthe oryzae). Twenty-seven PIBPs (PigmR-Interacting and Blast Resistance Protein) that interact with PigmR were screened. After repeated comparisons, the inventors predicted that PIBP1 has a strong interaction with PigmR.

[0233] In order to confirm t...

Embodiment 2

[0234] Example 2. PIBP1 positively regulates PigmR-mediated disease resistance but has no effect on Pish

[0235] In order to explore whether PIBP1 affects the resistance of PigmR to rice blast, the inventors constructed the PIBP1-RNAi and PIBP1-OE (PIBP1-GFP) in the context of NIL-Pigm (a near-isogenic line of Pigm in the Nipponbare background). transgenic line ( Figure 9 A and 9B) (PIBP1-RNAi / NIL-Pigm and pUBI::PIBP1-GFP(PIBP1-OE) / Nipponbare). Real-time results showed that PIBP1-RNAi or PIBP1-OE did not affect the expression of PigmR RNA ( Figure 9 C). Field injection of Magnaporthe grisea found that the PIBP1-RNAi strain was significantly susceptible to the disease compared with the control NIL-Pigm, while the overexpression strain had no difference from the NIL-Pigm ( figure 2 A). At the same time, the inventors obtained a PIBP1 knockout mutant (PIBP1CRISPR / Cas9(PIBP1-KO / NIL-Pigm)) in the NIL-Pigm background by using Crisper-Cas9 technology. Pathogen inoculation an...

Embodiment 3

[0237] Example 3, PIBP1 specifically interacts with broad-spectrum disease resistance genes and positively regulates Pizt-mediated disease resistance

[0238] In order to further explore whether PIBP1 is specifically involved in the immune process of PigmR, the inventors detected the interaction between PIBP1 and other NLRs. It was found that PIBP1 interacted with the broad-spectrum disease-resistant NLR genes Pizt and Pi9, but did not interact with the race-transforming NLR gene Pid3 ( figure 2 B, 2C, 9K and 9L). By protein sequence comparison, it was shown that this kind of broad-spectrum disease-resistant NLR protein is highly conserved in protein sequence ( Figure 9 M). It is suggested that PIBP1 may participate in the broad-spectrum disease resistance of plants by interacting with such conserved R genes. Subsequently, the inventor knocked out PIBP1 in the background of rice variety ZH11 (containing Pizt), and analyzed the pathogenic bacteria inoculation of the transg...

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Abstract

The invention relates to a transcription factor for enhancing broad-spectrum disease resistance of plants and an application. The invention discloses that PIBP1 directly interacts with PigmR and otherbroad-spectrum anti-disease R genes to promote accumulation of own cell nuclear proteins and positively regulate and control disease resistance of plants. The PIBP1 and the homologous gene are combined as a kind of non-classical transcription factors and activate expression of defensive genes such as OsWAK14 and OsPAL1 at the downstream of the non-classical transcription factors. Therefore, the invention provides a new transcription factor which activates a downstream disease-resistant gene through direct interaction with the R gene and endows plants with a new mechanism of disease resistance. The invention not only provides a practical and effective plant improvement method, but also provides a new way for broad-spectrum immune mechanism research of plants and disease resistance breedingof plants.

Description

technical field [0001] The invention belongs to the field of molecular biology or botany, and more specifically, the invention relates to a transcription factor for enhancing broad-spectrum disease resistance of plants and its application. Background technique [0002] Plant diseases can cause a substantial reduction in crop production, or even extinction, seriously affecting global food security. As the main food product in the world, rice feeds nearly half of the population. However, due to the damage of various pathogenic microorganisms to rice in nature, its yield will be reduced to varying degrees every year. Among them, fungal diseases such as rice blast (caused by Magnaporthe oryzae) and bacterial diseases such as bacterial blight (caused by Xanthomonas oryzae pv. oryzae) have the greatest impact on rice yield and quality serious. The annual production reduction caused by rice blast (Magnaporthe oryzae) alone is equivalent to 10-30% of the total production, which c...

Claims

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Application Information

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IPC IPC(8): C12N15/82C12N15/29C07K14/415C07K19/00
CPCC12N15/8279C07K14/415C07K2319/00
Inventor 何祖华翟科然邓一文李群
Owner CAS CENT FOR EXCELLENCE IN MOLECULAR PLANT SCI
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