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Novel gene necessary for production of conidium from Magnaporthe grisea and its uses

A conidia, piriformis technology, applied in genetic engineering, plant genetic improvement, application and other directions

Inactive Publication Date: 2009-07-01
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] From the above research reports, it can be seen that the formation and morphological construction of pyrosporium conidia is a very complicated process, and many genes may be involved in this process

Method used

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  • Novel gene necessary for production of conidium from Magnaporthe grisea and its uses
  • Novel gene necessary for production of conidium from Magnaporthe grisea and its uses
  • Novel gene necessary for production of conidium from Magnaporthe grisea and its uses

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Experimental program
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Effect test

Embodiment 1

[0039] Embodiment 1, pyrosporium conidia produce the isolation of essential gene MgCON1

[0040] 1. Screening and identification of mutants

[0041] a, the construction of the REMI transformant library of pyrosporium strain P131

[0042] A large amount of pUCATPH was extracted by alkaline lysis (Lu, S., Lyngholm, L., Yang, G., Bronson, C., Yoder, O.C. 1994. Tagged mutants at the Toxl locus of Cochliobolusheterostrophus by restriction enzyme-mediated integration. Proc. Natl.Acad.Sci.USA 91:1264-12653) of the plasmid DNA, respectively with restriction endonuclease HindIII, KpnI, SacI and SmaI digestion with its linearization, according to the method of REMI transformation (Shi Z., Christian D ., & Leung H., Enhanced transformation in Magnaporthe grisea by restriction enzyme mediated integration of plasmad DNA. Phytopathology, 1995, 85: 329-333.) Transform wild-type pyrosporium strain P131. Add 2 micrograms of linearized plasmid and corresponding restriction endonucleases to ea...

Embodiment 2

[0061] Demonstration of the role in the production of embodiment 2, MgCON1 pyrosporium conidia

[0062] Including complementation experiments and gene replacement experiments. This part includes the construction of complementary vectors and gene replacement vectors and the introduction of the above two types of vectors into Pyrosporium to obtain corresponding transformants. The construction of the complementary vector refers to the DNA fragment comprising the full-length functional sequence of MgCON1 (the 2253rd nucleotide to the 4019th nucleotide of contig2.767 in the Pyrosporium genome database) and a DNA fragment with a new The carrier of the mycin resistance gene was linked. The recipient strain of Pyricularia sp. introduced by the complementary vector is a mutant strain. Here, the neomycin resistance gene is not a limitation of the present invention, and other genes that can cause antibiotic resistance, that is, genes that cause fungal resistance other than the neomycin...

Embodiment 3

[0082] Embodiment 3, comparison of the infection ability of wild-type bacteria P131 and MgCON1 deletion mutant

[0083] The wild-type bacteria P131, MgCON1 deletion mutants mgcon1-1 and mgcon1-3 were inoculated on the tomato juice oatmeal medium plate, cultured at 26°C-28°C for 5 days, and fresh mycelium pieces of 3mm×3mm were cut with a scalpel As the inoculum adhered to the leaves of Lijiang Xintuan Heigu, the inoculated rice leaves were placed at 26°C in the dark for 24 hours, and then transferred to 26°C under light for 72 hours before taking pictures. From the results in Figure 7, it can be seen that the wild-type strain P131 formed typical susceptible lesions on rice leaves, while the MgCON1 deletion mutants mgcon1-1 and mgcon1-3 did not form susceptible lesions, indicating that it completely lost the ability to invade. The ability to dye rice (results are shown in Figure 7).

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Abstract

The invention provides a new gene MgCON1 necessary for the production of pyrosporium conidia and its use. The gene and its cDNA, encoded protein and promoter respectively have the nucleotide or amino acid sequences of SEQ ID No.: 1, No.: 2, No.: 3 and No.: 4 in the sequence table. The encoded product of this gene has a C2H2 zinc finger domain, is localized in the nucleus, and is specifically expressed in conidiophores, conidiophores, and appresses; its knockout results in complete inability to produce conidia in Pyrosporium and loss of ability to infect rice leaves. The expression of MgCON1, the cleavage of transcripts and the expression, modification and localization of the encoded protein, as well as the signaling pathways involved in the gene can be used as important candidate targets for the design and screening of new antifungal agents.

Description

technical field [0001] The invention relates to the application of the coding gene for controlling fungi to produce conidia and related proteins in the field of microbial genetic engineering and plant protection. Background technique [0002] Magnaporthe grisea is a fungus belonging to the subphylum Ascomycota, which can infect rice, wheat, barley, millet and other grasses, causing blast. In particular, the rice blast caused by the fungus infecting rice occurs every year in various rice cultivation areas in the world, and the damage is extensive and serious. In general, rice blast damage can reduce rice production by 5-10%, and severely diseased fields can lead to failure of rice harvest. Rice blast has been popular in my country for many times, and it is also one of the main diseases of rice in my country. [0003] The conidia of Pyrosporium are pear-shaped and consist of three cells. Usually 3 to 5 conidia are produced in a symaxial manner from the tip of a conidiophore...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/37C12N15/31C12N15/63A01N63/00
Inventor 彭友良张裕君赵文生
Owner CHINA AGRI UNIV
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