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Cochliobolus victoriae gene CvCAO1 and application thereof

A blight fungus and oat technology, which is applied in the field of discovery and the application of encoded proteins, can solve the problems of difficulty in preventing and controlling oat leaf blight, unclear production mechanism of toxin victorin, loss of oat, etc.

Active Publication Date: 2020-02-28
JILIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Oat leaf blight has caused great losses to oat production every year, especially the production of toxin victorin has a great impact on human and animal safety
The production mechanism of the toxin victorin is still unclear, which has caused great difficulties in the control of oat leaf blight

Method used

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  • Cochliobolus victoriae gene CvCAO1 and application thereof
  • Cochliobolus victoriae gene CvCAO1 and application thereof
  • Cochliobolus victoriae gene CvCAO1 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1 Correlation analysis of CvCAO1 gene

[0026] The CvCAO1 gene was obtained by analyzing the genome of Cochliobolus victoriae FI3. The open reading frame of the CvCAO1 gene of Leaf blight of oats consists of 2332 nucleotides, including 4 introns. The encoded protein product consists of 709 amino acids, and domain analysis revealed that the CvCAO1 protein contains a conserved Cu_amine_oxid functional domain (see figure 1 ).

Embodiment 2

[0027] The knockout of embodiment 2CvCAO1 gene

[0028] 1) The upstream and downstream of CvCAO1 gene and the amplification of hygromycin gene

[0029] Primers F1 (5'-CTAGAGATGAAGGCCCTGGA-3') and R1 (5'-TCCTGTGTGAAATTGTTATCCGCTAGGGTCCGAGTTCACACAAA-3') were used to amplify the 621bp fragment upstream of the CvCAO1 gene using F2(5 '-GTCGTGACTGGGAAAACCCTGGCGGAGAAGGAGTGCAGGTTTGG-3') and R2 (5'-GGTTTTCGCGGATGAAGTAA-3') to amplify the 816bp fragment downstream of the CvCAO1 gene of Leaf blight of oats 3'), using the vector pUCATPH as a template to amplify the 2549bp hygromycin gene. The reaction system is: 10mmol / L dNTP Mixture, 1μL; 5×PCR buffer, 10μL; each 2.5μL of upstream and downstream primers (10μmol / mL); template DNA, 2μL; Phusion polymerase, 0.5μL (5U); ddH 2 O, 31.5 μL; amplification program: 98°C pre-denaturation for 2 minutes, then (1) 98°C, denaturation for 20 seconds; (2) 65°C, annealing for 30 seconds; (3) 72°C, extension for 30 seconds; (4) ) cycled 30 times; (5) e...

Embodiment 3

[0037] Example 3 The role of CvCAO1 gene in the production of oat leaf blight (Cochliobolus victoriae) FI3 toxin

[0038] use

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Abstract

The invention belongs to the technical field of microbial gene engineering and discloses cochliobolus victoriae gene CvCAO1 and application thereof. A DNA sequence of the CvCAO1 gene derived from cochliobolus victoriae and controlling toxin formation is as shown in SEQ ID No:1; an amino sequence of protein coded by the CvCAO1 gene is as shown in SEQ ID No:2. The CvCAO1 gene can be applied to the field of plant anti-cochliobolus victoriae gene engineering. By deficiency, mutation or modification of toxin formation control protein CvCAO1 of cochliobolus victoriae, toxin limitation is realized, and application of the cochliobolus victoriae gene CvCAO1 as a target to design and scanning of anti-cochliobolus victoriae agents can be realized.

Description

technical field [0001] The invention belongs to the technical field of microbial genetic engineering, and specifically relates to the discovery of genes controlling the formation of mycotoxins in the field of plant protection and the application of encoded proteins. Background technique [0002] Oat leaf blight is caused by a fungus called Cochliobolus victoriae. Oat leaf blight mainly damages leaves, leaf sheaths, glumes and other shoots. In the early stage of leaf infection, yellow to yellow-brown lesions with inconspicuous edges will form, and then gradually expand, brown in the middle, and slightly yellow around. When there are many lesions, they gradually die down from the leaf tip. The disease mostly starts from the lower leaves and spreads upwards. Moreover, Oat Leaf Blight can produce a toxin-victorin, which can still be toxic to specific genotype oat varieties after being diluted one million times. Since then, the concept of host-specific toxin has been proposed a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/53C12N9/06
CPCC12N9/0022C12Y104/03006
Inventor 张祥辉于汇琳王璐孙颖焦文莉刘金亮潘洪玉
Owner JILIN UNIV