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Written | Qingfeng Mingyue Editor | Wang Yi In the plant immune activities, the disease-related protein-mediated immune process (PTI) and the effector-induced plant immune process (ETI) represent two levels of the interaction process between pathogens and host plants, respectively.
There are both differences and close ties between the people [1].
Pathogen-related molecular patterns (PAMPs) secreted by pathogens induce immunity by specifically recognizing pattern recognition receptors (PRRs) on the surface of host cells.
For plant pathogenic fungi, chitin oligosaccharides are typical PAMP [2,3].
Chitin is composed of a series of 1,4-linked N-acetylglucosamine, which is widely distributed in fungal cell walls, arthropod exoskeletons and nematodes [4,5].
During the process of plant pathogenic fungi infecting the host, the hydrolase produced by the host, such as chitinase, can degrade the fungal cell wall to produce chitin oligomers, which then activate the host immune response.
The molecules that recognize chitin oligomers are located on the cell surface.
CEBiP and CERK1 are known.
After these PPRs recognize chitin oligomers, they will induce a series of immune responses, including ROS accumulation and callose deposition [6, 7].
Cunning phytopathogenic fungi possess a special chitinase deacetylase inhibitory protein, which modifies the fungal cell membrane, thereby reducing the activity of degradation by the host chitinase and achieving the purpose of pathogen infection [8] .
Effectors with similar functions have been reported in Magnaporthe grisea and Sphaeropsis.
However, proteins with similar functions in powdery mildew have not been systematically studied.Recently, researchers from the National University of Malaga in Spain published a research paper entitled Effects With Chintinase Activity (EWCAs), a family of conserved, secreted fungal chitinases that suppress chitin-triggered immunity in The Plant Cell.
The study used gene expression analysis, RNAi silencing analysis, protein modeling and protein ligand binding prediction, enzyme activity detection and protein localization methods to deeply explore the effectors of EWCA (Effectors With Chitinase Activity) in cucurbit powdery mildew bacteria to inhibit plant immunity Mechanisms.
The study first identified a class of protein PECs with unknown functions in powdery mildew bacteria through sequence similarity analysis and detection of gene expression differences during infection; after silencing candidate PEC genes, the fungal biomass was evaluated by qPCR and mutants were found The colony becomes smaller and there are fewer haustoriums.
The corpus callosin deposition and hydrogen peroxide accumulation were further tested, and it was found that the mutant strain significantly induced the host's immunity compared with the wild type.
In order to explore the function of these proteins, the study used I-TASSER to model the PEC protein, and found that these proteins have a conserved area of unknown function (DUF).
Further bioinformatics analysis of potential receptor binding sites revealed that PEC Proteins are similar to the glycoside hydrolase family and may function as chitinases.
In order to study the chitinase activity of PEC protein, four PEC proteins were expressed in vitro to detect their binding ability to cellulose, colloidal chitin and xylan.
Biochemical experiments of different combinations confirmed the chitin of this protein family.
Chitinase activity, and named EWCAs (Effector With Chitinase Activity). The researchers made up for the reduced virulence phenotype of PECs mutants by simultaneously mutating PECs and the chitin receptor protein CERK1, and determined that EWCAs are involved in chitin-mediated immunity.
These were detected by fusion of GFP and functional expression of different EWCAs.
The protein is located in the papillae of the plant infected with the fungal haustorium.
The corpus callosum deposition experiment proved that the location of plant papillae is related to the effector function, and the confocal scanning analysis EWCA protein is always located in the small suction.
Finally, the researchers analyzed the conservation of EWCA protein in fungi and found that this type of protein is widely distributed in pathogenic fungi such as insects, nematodes, animals, etc.
, and they are all related to the DUF domain.
At the same time, they found that homologous genes of different species exist Multi-copy phenomenon.
To sum up, the study systematically analyzed a type of protein in cucurbit powdery mildew, and comprehensively used bioinformatics, biochemistry, genetics and other methods to confirm that EWCAs are a type of chitinase effector.
It mainly acts on the plant immunity induced by chitin, and is widely distributed in different fungi.
It deepens our understanding of the diversity of host and pathogen infection methods by plant pathogenic fungi, and provides a reference for us to further study the interaction between host and pathogenic fungi. References: [1] Jones, JD, and Dangl, JL (2006).
The plant immune system.
Nature 444, 323–329.
[2] Young, VL, Simpson, RM, and Ward, VK.
(2005).
Characterization of an exochitinase from Epiphyas postvittana nucleopolyhedrovirus (family Baculoviridae).
J Gen Virol 86, 3253-3261【3】de Jonge, R, van Esse, HP, Kombrink, A, Shinya, T, Desaki, Y, Bours, R, van der Krol, S,Shibuya, N, Joosten, MHAJ, and Thomma, BPHJ.
(2010).
Conserved fungal LysM effector Ecp6 prevents chitin-triggered immunity in plants.
Science 329, 953-955【4】Krokeide, IM, Synstad , B, Gåseidnes, S, Horn, SJ, Eijsink, VG, and Sørlie, M.
(2007).
Natural substrate assay for chitinase using high-performance liquid chromatography: a comparison with existing assays.
Anal Biochem 363, 128-134【 5] de Jonge, R, van Esse, HP, Kombrink, A, Shinya, T, Desaki, Y, Bours, R, van der Krol, S, Shibuya, N,Joosten, MHAJ, and Thomma, BPHJ.
(2010).
Conserved fungal LysM effector Ecp6 prevents chitin-triggered immunity in plants.
Science 329, 953-955 [6] van den Burg, HA, Harrison, SJ, Joosten, MH, Vervoort , J, and de Wit, PJ.
(2006).
Cladosporium fulvum Avr4 protects fungal cell walls against hydrolysis by plant chitinases accumulation during infection.
Mol Plant Microbe Interact 19, 1420-1430【7】Mentlak, TA, Kombrink, A, Shinya , T, Ryder, LS, Otomo, I, Saitoh, H, Terauchi, R, Nishizawa, Y, Shibuya, N, Thomma, BPHJ, and Talbot, NJ.
(2012).
Effector-mediated suppression of chitin-triggered immunity by Magnaporthe oryzae is necessary for rice blast disease.
Plant Cell 24, 322-335.
【8】Sánchez-Vallet, A, Saleem-Batcha, R, Kombrink, A, Hansen, G, Valkenburg, DJ, Thomma, BP, and Mesters , JR.
(2013).
Fungal effector Ecp6 outcompetes host immune receptor for chitin binding through intrachain LysM dimerization.
Elife 2, e00790 Original link: https://academic.
oup.
com/plcell/advance-article/doi/10.
1093/plcell/koab011/6119333
There are both differences and close ties between the people [1].
Pathogen-related molecular patterns (PAMPs) secreted by pathogens induce immunity by specifically recognizing pattern recognition receptors (PRRs) on the surface of host cells.
For plant pathogenic fungi, chitin oligosaccharides are typical PAMP [2,3].
Chitin is composed of a series of 1,4-linked N-acetylglucosamine, which is widely distributed in fungal cell walls, arthropod exoskeletons and nematodes [4,5].
During the process of plant pathogenic fungi infecting the host, the hydrolase produced by the host, such as chitinase, can degrade the fungal cell wall to produce chitin oligomers, which then activate the host immune response.
The molecules that recognize chitin oligomers are located on the cell surface.
CEBiP and CERK1 are known.
After these PPRs recognize chitin oligomers, they will induce a series of immune responses, including ROS accumulation and callose deposition [6, 7].
Cunning phytopathogenic fungi possess a special chitinase deacetylase inhibitory protein, which modifies the fungal cell membrane, thereby reducing the activity of degradation by the host chitinase and achieving the purpose of pathogen infection [8] .
Effectors with similar functions have been reported in Magnaporthe grisea and Sphaeropsis.
However, proteins with similar functions in powdery mildew have not been systematically studied.Recently, researchers from the National University of Malaga in Spain published a research paper entitled Effects With Chintinase Activity (EWCAs), a family of conserved, secreted fungal chitinases that suppress chitin-triggered immunity in The Plant Cell.
The study used gene expression analysis, RNAi silencing analysis, protein modeling and protein ligand binding prediction, enzyme activity detection and protein localization methods to deeply explore the effectors of EWCA (Effectors With Chitinase Activity) in cucurbit powdery mildew bacteria to inhibit plant immunity Mechanisms.
The study first identified a class of protein PECs with unknown functions in powdery mildew bacteria through sequence similarity analysis and detection of gene expression differences during infection; after silencing candidate PEC genes, the fungal biomass was evaluated by qPCR and mutants were found The colony becomes smaller and there are fewer haustoriums.
The corpus callosin deposition and hydrogen peroxide accumulation were further tested, and it was found that the mutant strain significantly induced the host's immunity compared with the wild type.
In order to explore the function of these proteins, the study used I-TASSER to model the PEC protein, and found that these proteins have a conserved area of unknown function (DUF).
Further bioinformatics analysis of potential receptor binding sites revealed that PEC Proteins are similar to the glycoside hydrolase family and may function as chitinases.
In order to study the chitinase activity of PEC protein, four PEC proteins were expressed in vitro to detect their binding ability to cellulose, colloidal chitin and xylan.
Biochemical experiments of different combinations confirmed the chitin of this protein family.
Chitinase activity, and named EWCAs (Effector With Chitinase Activity). The researchers made up for the reduced virulence phenotype of PECs mutants by simultaneously mutating PECs and the chitin receptor protein CERK1, and determined that EWCAs are involved in chitin-mediated immunity.
These were detected by fusion of GFP and functional expression of different EWCAs.
The protein is located in the papillae of the plant infected with the fungal haustorium.
The corpus callosum deposition experiment proved that the location of plant papillae is related to the effector function, and the confocal scanning analysis EWCA protein is always located in the small suction.
Finally, the researchers analyzed the conservation of EWCA protein in fungi and found that this type of protein is widely distributed in pathogenic fungi such as insects, nematodes, animals, etc.
, and they are all related to the DUF domain.
At the same time, they found that homologous genes of different species exist Multi-copy phenomenon.
To sum up, the study systematically analyzed a type of protein in cucurbit powdery mildew, and comprehensively used bioinformatics, biochemistry, genetics and other methods to confirm that EWCAs are a type of chitinase effector.
It mainly acts on the plant immunity induced by chitin, and is widely distributed in different fungi.
It deepens our understanding of the diversity of host and pathogen infection methods by plant pathogenic fungi, and provides a reference for us to further study the interaction between host and pathogenic fungi. References: [1] Jones, JD, and Dangl, JL (2006).
The plant immune system.
Nature 444, 323–329.
[2] Young, VL, Simpson, RM, and Ward, VK.
(2005).
Characterization of an exochitinase from Epiphyas postvittana nucleopolyhedrovirus (family Baculoviridae).
J Gen Virol 86, 3253-3261【3】de Jonge, R, van Esse, HP, Kombrink, A, Shinya, T, Desaki, Y, Bours, R, van der Krol, S,Shibuya, N, Joosten, MHAJ, and Thomma, BPHJ.
(2010).
Conserved fungal LysM effector Ecp6 prevents chitin-triggered immunity in plants.
Science 329, 953-955【4】Krokeide, IM, Synstad , B, Gåseidnes, S, Horn, SJ, Eijsink, VG, and Sørlie, M.
(2007).
Natural substrate assay for chitinase using high-performance liquid chromatography: a comparison with existing assays.
Anal Biochem 363, 128-134【 5] de Jonge, R, van Esse, HP, Kombrink, A, Shinya, T, Desaki, Y, Bours, R, van der Krol, S, Shibuya, N,Joosten, MHAJ, and Thomma, BPHJ.
(2010).
Conserved fungal LysM effector Ecp6 prevents chitin-triggered immunity in plants.
Science 329, 953-955 [6] van den Burg, HA, Harrison, SJ, Joosten, MH, Vervoort , J, and de Wit, PJ.
(2006).
Cladosporium fulvum Avr4 protects fungal cell walls against hydrolysis by plant chitinases accumulation during infection.
Mol Plant Microbe Interact 19, 1420-1430【7】Mentlak, TA, Kombrink, A, Shinya , T, Ryder, LS, Otomo, I, Saitoh, H, Terauchi, R, Nishizawa, Y, Shibuya, N, Thomma, BPHJ, and Talbot, NJ.
(2012).
Effector-mediated suppression of chitin-triggered immunity by Magnaporthe oryzae is necessary for rice blast disease.
Plant Cell 24, 322-335.
【8】Sánchez-Vallet, A, Saleem-Batcha, R, Kombrink, A, Hansen, G, Valkenburg, DJ, Thomma, BP, and Mesters , JR.
(2013).
Fungal effector Ecp6 outcompetes host immune receptor for chitin binding through intrachain LysM dimerization.
Elife 2, e00790 Original link: https://academic.
oup.
com/plcell/advance-article/doi/10.
1093/plcell/koab011/6119333
