Assessing European wheat sensitivities to Parastagonospora nodorum necrotrophic effectors and fine-mapping the Snn3-B1 locus conferring sensitivity to the effector SnTox3

Rowena Downie, Laura Bouvet, Eiko Furuki, Nick Gosman, Keith Gardner, Ian Mackay, Camila Campos Mantello, Greg Mellers, Huyen Phan, Gemma Rose, Kar-Chun Tan, Richard Oliver, James Cockram

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Here, we evaluate the expression of the proteinaceous effectors ToxA and ToxB, produced by the necrotrophic fungal pathogen Pyrenophora tritici-repentis, which confer tan spot disease susceptibility on wheat. These necrotrophic effectors were expressed in two heterologous systems: Escherichia coli and Pichia pastoris. The E. coli SHuffle system was demonstrated to be superior to P. pastoris in generating high-levels of recombinant proteins that were soluble and stable. In addition, protein extracts from P. pastoris induced non-specific chlorosis on wheat, postulated to be caused by co-purified glucanases secreted by the host. Up to 79.6 μg/ml of ToxB was obtained using the SHuffle system in the absence of the native signal peptide, whilst the ToxA yield was considerably lower at 3.2 μg/ml. Results indicated that a histidine tag at the ToxA C-terminus interfered with effector functionality. Heterologously expressed ToxA and ToxB were tested on a panel of Australian cereals, including 122 varieties of bread wheat, 16 durum, 20 triticale and 5 barley varieties, as well as common plant model species including tobacco and Arabidopsis thaliana. A varying degree of effector sensitivities was observed, with a higher ToxB sensitivity and prevalence in the durum and triticale varieties. ToxB-induced chlorosis was also detected on barley. The heterologous expression of effectors that are easily scalable, will facilitate effector-assisted selection of varieties in wheat breeding programs as well as the investigation of P. tritici-repentis effectors in host and non-host interactions.
Original languageEnglish
Number of pages12
JournalFrontiers in Plant Science
Volume9:881
Early online date4 Jul 2018
DOIs
Publication statusFirst published - 4 Jul 2018
Externally publishedYes

Fingerprint

Pichia pastoris
Triticum
Hypochromic Anemia
loci
wheat
triticale
Hordeum
chlorosis
durum wheat
barley
Pyrenophora tritici-repentis
Escherichia coli
Pichia
Bread
Disease Susceptibility
Protein Sorting Signals
signal peptide
histidine
Recombinant Proteins
Arabidopsis

Keywords

  • wheat
  • MAGIC
  • Plant breeding
  • Plant disease

Cite this

Downie, Rowena ; Bouvet, Laura ; Furuki, Eiko ; Gosman, Nick ; Gardner, Keith ; Mackay, Ian ; Campos Mantello, Camila ; Mellers, Greg ; Phan, Huyen ; Rose, Gemma ; Tan, Kar-Chun ; Oliver, Richard ; Cockram, James. / Assessing European wheat sensitivities to Parastagonospora nodorum necrotrophic effectors and fine-mapping the Snn3-B1 locus conferring sensitivity to the effector SnTox3. In: Frontiers in Plant Science. 2018 ; Vol. 9:881.
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abstract = "Here, we evaluate the expression of the proteinaceous effectors ToxA and ToxB, produced by the necrotrophic fungal pathogen Pyrenophora tritici-repentis, which confer tan spot disease susceptibility on wheat. These necrotrophic effectors were expressed in two heterologous systems: Escherichia coli and Pichia pastoris. The E. coli SHuffle system was demonstrated to be superior to P. pastoris in generating high-levels of recombinant proteins that were soluble and stable. In addition, protein extracts from P. pastoris induced non-specific chlorosis on wheat, postulated to be caused by co-purified glucanases secreted by the host. Up to 79.6 μg/ml of ToxB was obtained using the SHuffle system in the absence of the native signal peptide, whilst the ToxA yield was considerably lower at 3.2 μg/ml. Results indicated that a histidine tag at the ToxA C-terminus interfered with effector functionality. Heterologously expressed ToxA and ToxB were tested on a panel of Australian cereals, including 122 varieties of bread wheat, 16 durum, 20 triticale and 5 barley varieties, as well as common plant model species including tobacco and Arabidopsis thaliana. A varying degree of effector sensitivities was observed, with a higher ToxB sensitivity and prevalence in the durum and triticale varieties. ToxB-induced chlorosis was also detected on barley. The heterologous expression of effectors that are easily scalable, will facilitate effector-assisted selection of varieties in wheat breeding programs as well as the investigation of P. tritici-repentis effectors in host and non-host interactions.",
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Assessing European wheat sensitivities to Parastagonospora nodorum necrotrophic effectors and fine-mapping the Snn3-B1 locus conferring sensitivity to the effector SnTox3. / Downie, Rowena; Bouvet, Laura; Furuki, Eiko; Gosman, Nick; Gardner, Keith; Mackay, Ian; Campos Mantello, Camila; Mellers, Greg; Phan, Huyen; Rose, Gemma; Tan, Kar-Chun; Oliver, Richard; Cockram, James.

In: Frontiers in Plant Science, Vol. 9:881, 04.07.2018.

Research output: Contribution to journalArticle

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AU - Downie, Rowena

AU - Bouvet, Laura

AU - Furuki, Eiko

AU - Gosman, Nick

AU - Gardner, Keith

AU - Mackay, Ian

AU - Campos Mantello, Camila

AU - Mellers, Greg

AU - Phan, Huyen

AU - Rose, Gemma

AU - Tan, Kar-Chun

AU - Oliver, Richard

AU - Cockram, James

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AB - Here, we evaluate the expression of the proteinaceous effectors ToxA and ToxB, produced by the necrotrophic fungal pathogen Pyrenophora tritici-repentis, which confer tan spot disease susceptibility on wheat. These necrotrophic effectors were expressed in two heterologous systems: Escherichia coli and Pichia pastoris. The E. coli SHuffle system was demonstrated to be superior to P. pastoris in generating high-levels of recombinant proteins that were soluble and stable. In addition, protein extracts from P. pastoris induced non-specific chlorosis on wheat, postulated to be caused by co-purified glucanases secreted by the host. Up to 79.6 μg/ml of ToxB was obtained using the SHuffle system in the absence of the native signal peptide, whilst the ToxA yield was considerably lower at 3.2 μg/ml. Results indicated that a histidine tag at the ToxA C-terminus interfered with effector functionality. Heterologously expressed ToxA and ToxB were tested on a panel of Australian cereals, including 122 varieties of bread wheat, 16 durum, 20 triticale and 5 barley varieties, as well as common plant model species including tobacco and Arabidopsis thaliana. A varying degree of effector sensitivities was observed, with a higher ToxB sensitivity and prevalence in the durum and triticale varieties. ToxB-induced chlorosis was also detected on barley. The heterologous expression of effectors that are easily scalable, will facilitate effector-assisted selection of varieties in wheat breeding programs as well as the investigation of P. tritici-repentis effectors in host and non-host interactions.

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KW - MAGIC

KW - Plant breeding

KW - Plant disease

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