A family of small tyrosine rich proteins is essential for oogonial and oospore cell wall development of the mycoparasitic oomycete Pythium oligandrum

Laura J. Grenville-Briggs, Neil R. Horner, Andrew J. Phillips, Gordon W. Beakes, Pieter Van West*

*Corresponding author for this work

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

The mycoparasitic oomycete Pythium oligandrum is homothallic, producing an abundance of thick-walled spiny oospores in culture. After mining a cDNA sequence dataset, we identified a family of genes that code for small tyrosine rich (Pythium oligandrum small tyrosine rich (PoStr)) proteins. Sequence analysis identified similarity between the PoStr proteins and putative glycine-rich cell wall proteins from the related plant pathogenic oomycete Pythium ultimum, and mating-induced genes from the oomycete Phytophthora infestans. Expression analysis showed that PoStr transcripts accumulate during oospore production in culture and immunolocalisation indicates the presence of these proteins in oogonial and oospore cell walls. PoStr protein abundance correlated positively with production of oogonia as determined by antibiotic-mediated oogonia suppression. To further characterise the role of PoStr proteins in P. oligandrum oospore production, we silenced this gene family using homology-dependent gene silencing. This represents the first characterisation of genes using gene silencing in a Pythium species. Oospores from silenced strains displayed major ultrastructural changes and were sensitive to degradative enzyme treatment. Oogonia of silenced strains either appeared to be arrested at the mature oosphere stage of development or in around 40 % of the structures, showed a complete suppression of oospore formation. Suppressed oogonia were highly vacuolated and the oogonium wall was thickened by a new inner wall layer. Our data suggest PoStr proteins are probably integral structural components of the normal oospore cell wall and play a key role in oospore formation. (C) 2013 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

Original languageEnglish
Pages (from-to)163-172
Number of pages10
JournalFungal Biology
Volume117
Issue number3
Early online date19 Jan 2013
DOIs
Publication statusPublished - Mar 2013

Keywords

  • gene Silencing
  • mycoparasite
  • oomycete
  • oomycete sexual reproduction
  • oospore
  • oospore formation
  • oosporogenesis
  • pythium
  • TEM
  • ultrastructure
  • phytophthora-infestans
  • appressorium formation
  • damping-off
  • infection
  • gene
  • ultimum
  • aphanidermatum
  • transformation
  • germination

Cite this

A family of small tyrosine rich proteins is essential for oogonial and oospore cell wall development of the mycoparasitic oomycete Pythium oligandrum. / Grenville-Briggs, Laura J.; Horner, Neil R.; Phillips, Andrew J.; Beakes, Gordon W.; Van West, Pieter.

In: Fungal Biology, Vol. 117, No. 3, 03.2013, p. 163-172.

Research output: Contribution to journalArticle

Grenville-Briggs, Laura J. ; Horner, Neil R. ; Phillips, Andrew J. ; Beakes, Gordon W. ; Van West, Pieter. / A family of small tyrosine rich proteins is essential for oogonial and oospore cell wall development of the mycoparasitic oomycete Pythium oligandrum. In: Fungal Biology. 2013 ; Vol. 117, No. 3. pp. 163-172.
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title = "A family of small tyrosine rich proteins is essential for oogonial and oospore cell wall development of the mycoparasitic oomycete Pythium oligandrum",
abstract = "The mycoparasitic oomycete Pythium oligandrum is homothallic, producing an abundance of thick-walled spiny oospores in culture. After mining a cDNA sequence dataset, we identified a family of genes that code for small tyrosine rich (Pythium oligandrum small tyrosine rich (PoStr)) proteins. Sequence analysis identified similarity between the PoStr proteins and putative glycine-rich cell wall proteins from the related plant pathogenic oomycete Pythium ultimum, and mating-induced genes from the oomycete Phytophthora infestans. Expression analysis showed that PoStr transcripts accumulate during oospore production in culture and immunolocalisation indicates the presence of these proteins in oogonial and oospore cell walls. PoStr protein abundance correlated positively with production of oogonia as determined by antibiotic-mediated oogonia suppression. To further characterise the role of PoStr proteins in P. oligandrum oospore production, we silenced this gene family using homology-dependent gene silencing. This represents the first characterisation of genes using gene silencing in a Pythium species. Oospores from silenced strains displayed major ultrastructural changes and were sensitive to degradative enzyme treatment. Oogonia of silenced strains either appeared to be arrested at the mature oosphere stage of development or in around 40 {\%} of the structures, showed a complete suppression of oospore formation. Suppressed oogonia were highly vacuolated and the oogonium wall was thickened by a new inner wall layer. Our data suggest PoStr proteins are probably integral structural components of the normal oospore cell wall and play a key role in oospore formation. (C) 2013 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.",
keywords = "gene Silencing, mycoparasite, oomycete, oomycete sexual reproduction, oospore, oospore formation, oosporogenesis, pythium, TEM, ultrastructure, phytophthora-infestans, appressorium formation, damping-off, infection, gene, ultimum, aphanidermatum, transformation, germination",
author = "Grenville-Briggs, {Laura J.} and Horner, {Neil R.} and Phillips, {Andrew J.} and Beakes, {Gordon W.} and {Van West}, Pieter",
note = "Our work is supported by the University of Aberdeen (PvW), the Biotechnology and Biological Sciences Research Council (BBSRC) (LGB, PvW), the Royal Society (PvW), the Total Foundation (PvW), the EU (LGB) via a Marie Curie Intra European Fellowship (FP7-PEOPLE-2010-IEF; CBOP) and the Natural Environment Research Council (NERC) via the Strategic Ocean Funding Initiative Initiative (award NE/F012705/1) (LGB, PvW) and an NERC PhD-studentship (NH). Funding sources had no role in experimental design, analysis, interpretation of data, or in the preparation of this manuscript. Plasmid pTOR was a gift from Felix Mauch.",
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T1 - A family of small tyrosine rich proteins is essential for oogonial and oospore cell wall development of the mycoparasitic oomycete Pythium oligandrum

AU - Grenville-Briggs, Laura J.

AU - Horner, Neil R.

AU - Phillips, Andrew J.

AU - Beakes, Gordon W.

AU - Van West, Pieter

N1 - Our work is supported by the University of Aberdeen (PvW), the Biotechnology and Biological Sciences Research Council (BBSRC) (LGB, PvW), the Royal Society (PvW), the Total Foundation (PvW), the EU (LGB) via a Marie Curie Intra European Fellowship (FP7-PEOPLE-2010-IEF; CBOP) and the Natural Environment Research Council (NERC) via the Strategic Ocean Funding Initiative Initiative (award NE/F012705/1) (LGB, PvW) and an NERC PhD-studentship (NH). Funding sources had no role in experimental design, analysis, interpretation of data, or in the preparation of this manuscript. Plasmid pTOR was a gift from Felix Mauch.

PY - 2013/3

Y1 - 2013/3

N2 - The mycoparasitic oomycete Pythium oligandrum is homothallic, producing an abundance of thick-walled spiny oospores in culture. After mining a cDNA sequence dataset, we identified a family of genes that code for small tyrosine rich (Pythium oligandrum small tyrosine rich (PoStr)) proteins. Sequence analysis identified similarity between the PoStr proteins and putative glycine-rich cell wall proteins from the related plant pathogenic oomycete Pythium ultimum, and mating-induced genes from the oomycete Phytophthora infestans. Expression analysis showed that PoStr transcripts accumulate during oospore production in culture and immunolocalisation indicates the presence of these proteins in oogonial and oospore cell walls. PoStr protein abundance correlated positively with production of oogonia as determined by antibiotic-mediated oogonia suppression. To further characterise the role of PoStr proteins in P. oligandrum oospore production, we silenced this gene family using homology-dependent gene silencing. This represents the first characterisation of genes using gene silencing in a Pythium species. Oospores from silenced strains displayed major ultrastructural changes and were sensitive to degradative enzyme treatment. Oogonia of silenced strains either appeared to be arrested at the mature oosphere stage of development or in around 40 % of the structures, showed a complete suppression of oospore formation. Suppressed oogonia were highly vacuolated and the oogonium wall was thickened by a new inner wall layer. Our data suggest PoStr proteins are probably integral structural components of the normal oospore cell wall and play a key role in oospore formation. (C) 2013 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

AB - The mycoparasitic oomycete Pythium oligandrum is homothallic, producing an abundance of thick-walled spiny oospores in culture. After mining a cDNA sequence dataset, we identified a family of genes that code for small tyrosine rich (Pythium oligandrum small tyrosine rich (PoStr)) proteins. Sequence analysis identified similarity between the PoStr proteins and putative glycine-rich cell wall proteins from the related plant pathogenic oomycete Pythium ultimum, and mating-induced genes from the oomycete Phytophthora infestans. Expression analysis showed that PoStr transcripts accumulate during oospore production in culture and immunolocalisation indicates the presence of these proteins in oogonial and oospore cell walls. PoStr protein abundance correlated positively with production of oogonia as determined by antibiotic-mediated oogonia suppression. To further characterise the role of PoStr proteins in P. oligandrum oospore production, we silenced this gene family using homology-dependent gene silencing. This represents the first characterisation of genes using gene silencing in a Pythium species. Oospores from silenced strains displayed major ultrastructural changes and were sensitive to degradative enzyme treatment. Oogonia of silenced strains either appeared to be arrested at the mature oosphere stage of development or in around 40 % of the structures, showed a complete suppression of oospore formation. Suppressed oogonia were highly vacuolated and the oogonium wall was thickened by a new inner wall layer. Our data suggest PoStr proteins are probably integral structural components of the normal oospore cell wall and play a key role in oospore formation. (C) 2013 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

KW - gene Silencing

KW - mycoparasite

KW - oomycete

KW - oomycete sexual reproduction

KW - oospore

KW - oospore formation

KW - oosporogenesis

KW - pythium

KW - TEM

KW - ultrastructure

KW - phytophthora-infestans

KW - appressorium formation

KW - damping-off

KW - infection

KW - gene

KW - ultimum

KW - aphanidermatum

KW - transformation

KW - germination

U2 - 10.1016/j.funbio.2013.01.001

DO - 10.1016/j.funbio.2013.01.001

M3 - Article

VL - 117

SP - 163

EP - 172

JO - Fungal Biology

JF - Fungal Biology

SN - 1878-6146

IS - 3

ER -