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Botrytis cinerea protein O-mannosyltransferases play critical roles in morphogenesis, growth, and virulence.

González M, Brito N, Frías M, González C - PLoS ONE (2013)

Bottom Line: The most significant case is that of grapevine leaves, whose penetration requires the three functional PMTs.Furthermore, PMT2 also contributes significantly to fungal adherence on grapevine and tobacco leaves.Analysis of extracellular and membrane proteins showed significant changes in the pattern of protein secretion and glycosylation by the pmt mutants, and allowed the identification of new protein substrates putatively glycosylated by specific PMTs.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Bioquímica y Biología Molecular, Universidad de La Laguna, La Laguna (Tenerife), Spain.

ABSTRACT
Protein O-glycosylation is crucial in determining the structure and function of numerous secreted and membrane-bound proteins. In fungi, this process begins with the addition of a mannose residue by protein O-mannosyltransferases (PMTs) in the lumen side of the ER membrane. We have generated mutants of the three Botrytis cinerea pmt genes to study their role in the virulence of this wide-range plant pathogen. B. cinerea PMTs, especially PMT2, are critical for the stability of the cell wall and are necessary for sporulation and for the generation of the extracellular matrix. PMTs are also individually required for full virulence in a variety of hosts, with a special role in the penetration of intact plant leaves. The most significant case is that of grapevine leaves, whose penetration requires the three functional PMTs. Furthermore, PMT2 also contributes significantly to fungal adherence on grapevine and tobacco leaves. Analysis of extracellular and membrane proteins showed significant changes in the pattern of protein secretion and glycosylation by the pmt mutants, and allowed the identification of new protein substrates putatively glycosylated by specific PMTs. Since plants do no possess these enzymes, PMTs constitute a promising target in the development of novel control strategies against B. cinerea.

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Related in: MedlinePlus

Δbcpmt2 is impaired in adhesion to plant surfaces.A) Grapevine and tobacco leaves, inoculated at random positions with mycelium plugs from the three Δbcpmt mutants and the wild type (B05.10), were washed with water after 24-hours growth to remove non-attached plugs. B) Adhesion strength (mean values and standard deviation, n≥10) of agar plugs on grapevine and tobacco leaves at different times (hpi: hours after inoculation). Asterisks indicate a statistically significant difference with the wild type in the same conditions (p<0.05).
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pone-0065924-g010: Δbcpmt2 is impaired in adhesion to plant surfaces.A) Grapevine and tobacco leaves, inoculated at random positions with mycelium plugs from the three Δbcpmt mutants and the wild type (B05.10), were washed with water after 24-hours growth to remove non-attached plugs. B) Adhesion strength (mean values and standard deviation, n≥10) of agar plugs on grapevine and tobacco leaves at different times (hpi: hours after inoculation). Asterisks indicate a statistically significant difference with the wild type in the same conditions (p<0.05).

Mentions: Another indirect test for the lack of penetration/adherence of Δbcpmt mutants on intact plant surfaces was carried out by measuring the strength with which mycelial plugs adhere to tobacco or grapevine leaves, at different times after inoculation. In first place, we tested the resistance of plugs to be washed off from the leaf surface by water, one day after inoculation and before any symptom could be seen in the leaves. On grapevine leaves, only the Δbcpmt2 mutant could be removed by gentle agitation with water, while on tobacco leaves all Δbcpmt2 and Δbcpmt4 plugs were removed, as well as part of the Δbcpmt1 plugs (Figure 10A). The plugs with the wild type remained in the leaves in all occasions. The determination of the adherence strength, by pulling the agar plugs with a dynamometer, confirmed the lack of adherence of the plugs with Δbcpmt2, and also showed that Δbcpmt1 and Δbcpmt4 tend to bind to the leaf surface with less strength than the wild type (Figure 10B).


Botrytis cinerea protein O-mannosyltransferases play critical roles in morphogenesis, growth, and virulence.

González M, Brito N, Frías M, González C - PLoS ONE (2013)

Δbcpmt2 is impaired in adhesion to plant surfaces.A) Grapevine and tobacco leaves, inoculated at random positions with mycelium plugs from the three Δbcpmt mutants and the wild type (B05.10), were washed with water after 24-hours growth to remove non-attached plugs. B) Adhesion strength (mean values and standard deviation, n≥10) of agar plugs on grapevine and tobacco leaves at different times (hpi: hours after inoculation). Asterisks indicate a statistically significant difference with the wild type in the same conditions (p<0.05).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3675079&req=5

pone-0065924-g010: Δbcpmt2 is impaired in adhesion to plant surfaces.A) Grapevine and tobacco leaves, inoculated at random positions with mycelium plugs from the three Δbcpmt mutants and the wild type (B05.10), were washed with water after 24-hours growth to remove non-attached plugs. B) Adhesion strength (mean values and standard deviation, n≥10) of agar plugs on grapevine and tobacco leaves at different times (hpi: hours after inoculation). Asterisks indicate a statistically significant difference with the wild type in the same conditions (p<0.05).
Mentions: Another indirect test for the lack of penetration/adherence of Δbcpmt mutants on intact plant surfaces was carried out by measuring the strength with which mycelial plugs adhere to tobacco or grapevine leaves, at different times after inoculation. In first place, we tested the resistance of plugs to be washed off from the leaf surface by water, one day after inoculation and before any symptom could be seen in the leaves. On grapevine leaves, only the Δbcpmt2 mutant could be removed by gentle agitation with water, while on tobacco leaves all Δbcpmt2 and Δbcpmt4 plugs were removed, as well as part of the Δbcpmt1 plugs (Figure 10A). The plugs with the wild type remained in the leaves in all occasions. The determination of the adherence strength, by pulling the agar plugs with a dynamometer, confirmed the lack of adherence of the plugs with Δbcpmt2, and also showed that Δbcpmt1 and Δbcpmt4 tend to bind to the leaf surface with less strength than the wild type (Figure 10B).

Bottom Line: The most significant case is that of grapevine leaves, whose penetration requires the three functional PMTs.Furthermore, PMT2 also contributes significantly to fungal adherence on grapevine and tobacco leaves.Analysis of extracellular and membrane proteins showed significant changes in the pattern of protein secretion and glycosylation by the pmt mutants, and allowed the identification of new protein substrates putatively glycosylated by specific PMTs.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Bioquímica y Biología Molecular, Universidad de La Laguna, La Laguna (Tenerife), Spain.

ABSTRACT
Protein O-glycosylation is crucial in determining the structure and function of numerous secreted and membrane-bound proteins. In fungi, this process begins with the addition of a mannose residue by protein O-mannosyltransferases (PMTs) in the lumen side of the ER membrane. We have generated mutants of the three Botrytis cinerea pmt genes to study their role in the virulence of this wide-range plant pathogen. B. cinerea PMTs, especially PMT2, are critical for the stability of the cell wall and are necessary for sporulation and for the generation of the extracellular matrix. PMTs are also individually required for full virulence in a variety of hosts, with a special role in the penetration of intact plant leaves. The most significant case is that of grapevine leaves, whose penetration requires the three functional PMTs. Furthermore, PMT2 also contributes significantly to fungal adherence on grapevine and tobacco leaves. Analysis of extracellular and membrane proteins showed significant changes in the pattern of protein secretion and glycosylation by the pmt mutants, and allowed the identification of new protein substrates putatively glycosylated by specific PMTs. Since plants do no possess these enzymes, PMTs constitute a promising target in the development of novel control strategies against B. cinerea.

Show MeSH
Related in: MedlinePlus