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Barley stripe pathogenic gene PgPBS and applications thereof

A pathogenic gene and pathogenic technology, applied in application, genetic engineering, plant genetic improvement, etc., can solve the problems of no PgPBS gene and reports on the application of silencing this gene.

Active Publication Date: 2019-04-12
GANSU AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Problems in the prior art: there is no report on the application of the PgPBS gene and the application of silencing the gene in the prior art

Method used

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  • Barley stripe pathogenic gene PgPBS and applications thereof
  • Barley stripe pathogenic gene PgPBS and applications thereof
  • Barley stripe pathogenic gene PgPBS and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0051] This embodiment provides a barley stripe disease pathogenic gene PgPBS, the gene sequence is shown in SEQ ID NO: 1 in the sequence listing.

Embodiment 2

[0053] This embodiment provides a method for identifying the causative gene of barley stripe disease, comprising the steps of:

[0054] 1. Cultivation of barley streak disease strain

[0055] Isolate mycelia (strain number QWC) from barley leaves collected in Qinwangchuan, Gansu, put the mycelia in PD liquid medium, culture them on a shaking table at 25-30°C, 180-200 rpm for 2-3 days, and collect the mycelia Cells for the preparation of DNA, RNA and protoplasts. The formula of PDA solid medium was 200 g / L potato, 20 g / L glucose, and 15 g / L agar. The formula of PD liquid medium is: 200 g / L potato, 20 g / L glucose.

[0056] 2. Cloning and sequence analysis of PgPBS gene

[0057] The DNA and RNA of mycelial cells were extracted using a DNA kit (Omega BiotekInc, Norcross, GA) and an RNA kit (OmegaBiotekInc), respectively. The above RNA was reverse-transcribed to synthesize cDNA using a reverse transcription kit (TaKaRa Biotech, Dalian, China). Attached Figure 5 The primers (...

Embodiment 3

[0072] The present embodiment provides the application of a barley stripe disease pathogenic gene PgPBS, specifically including:

[0073] Application of a barley stripe disease pathogenic gene PgPBS to reduce the permeability of barley stripe disease pathogenic bacteria.

[0074] for research PgPBS Adaptation in stress treatments, WT and mutant (△PgPbs1, △PgPbs31, △PgPbs7) strains were subjected to the following treatments: ionic stress (Na + ), oxidative stress (H 2 o 2 ), heavy metal stress (CoCl 2 ), osmotic stress (sorbitol) and fungicide stress (dicarboximide fungicides and triazole tebuconazole fungicides). The radial growth rates of △PgPbs1, △PgPbs31 and △PgPbs7 under sorbitol stress were 8.6, 9.4 and 11.4 mm / d, respectively, and the radial growth rates under NaCl stress were 2.7, 3.1 and 5.0 mm / d, respectively, while the QWC The radial growth rates were 11.0 and 5.57 mm / d. Compared with the WT strain, the growth rates of △PgPbs1 and △PgPbs31 were significantly i...

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Abstract

Barley stripe is a seed-dispersal disease caused by a fungal pathogen-pyrenophora graminea, and usually causes severe barley yield loss. The invention relates to a method for identifying a barley stripe pathogenic gene, amplifies a PgPBS gene based on a homologous cloning technique, obtains a PgPBS-silenced transformant by an RNA interference (RNAi) technique, and further obtains applications of the gene in osmotic reaction, vegetative differentiation, cell wall integrity, drug resistance and pathogenicity.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and specifically relates to the pathogenic gene PgPBS of barley stripe disease and its application in maintaining the osmotic balance of pathogenic bacteria cells, protecting the integrity of pathogenic bacteria cell walls, and silencing the gene to reduce the pathogenicity of barley stripe disease . Background technique [0002] Barley stripe disease is caused by the fungal pathogen Sclerotinia sativae ( Drechslera gramina [Rabenh. ex. Schlech.] Shoemaker), often causing severe reductions in barley yields. Its yield loss can be mitigated by the application of fungicides and breeding for disease resistance, which is often considered a more economical and environmentally friendly solution. Therefore, it is of great significance to understand the genetic basis of toxicity and pathogenicity of this disease. [0003] Two-component signaling (TCS) systems regulate cell motility, control cell...

Claims

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Application Information

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IPC IPC(8): C12N15/54C12N15/80C12N1/15
CPCC12N9/12C12N15/80C12Y207/12002
Inventor 姚立蓉王化俊汪军成梁倩倩李葆春孟亚雄马小乐杨轲王刚马增科侯静静胡娜
Owner GANSU AGRI UNIV
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