Show simple item record

dc.contributor.authorMoffat, Caroline
dc.contributor.authorSee, Pao Theen
dc.contributor.authorIagallo, Elyce M
dc.contributor.authorOliver, Richard P
dc.date.accessioned2019-04-09T02:30:23Z
dc.date.available2019-04-09T02:30:23Z
dc.date.issued2019
dc.identifier.citationSee, P.T. and Iagallo, E.M. and Oliver, R.P. and Moffat, C.S. 2019. Heterologous Expression of the Pyrenophora tritici-repentis Effector Proteins ToxA and ToxB, and the Prevalence of Effector Sensitivity in Australian Cereal Crops. Frontiers in Microbiology. 10: Article ID 182.
dc.identifier.urihttp://hdl.handle.net/20.500.11937/75213
dc.identifier.doi10.3389/fmicb.2019.00182
dc.description.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.

dc.languageEnglish
dc.publisherFRONTIERS MEDIA SA
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/
dc.subjectScience & Technology
dc.subjectLife Sciences & Biomedicine
dc.subjectMicrobiology
dc.subjectprotein expression
dc.subjecttan spot
dc.subjectPyrenophora tritici-repentis
dc.subjectnecrotrophic effector
dc.subjectToxA
dc.subjectToxB
dc.subjectwheat
dc.subjectyellow spot
dc.subjectHOST-SELECTIVE TOXINS
dc.subjectTAN SPOT
dc.subjectPTR TOXB
dc.subjectNECROTROPHIC EFFECTORS
dc.subjectESCHERICHIA-COLI
dc.subject5 RACES
dc.subjectWHEAT
dc.subjectGENE
dc.subjectNODORUM
dc.subjectLOCUS
dc.titleHeterologous Expression of the Pyrenophora tritici-repentis Effector Proteins ToxA and ToxB, and the Prevalence of Effector Sensitivity in Australian Cereal Crops
dc.typeJournal Article
dcterms.source.volume10
dcterms.source.issn1664-302X
dcterms.source.titleFrontiers in Microbiology
dc.date.updated2019-04-09T02:25:40Z
curtin.departmentSchool of Molecular and Life Sciences (MLS)
curtin.accessStatusOpen access
curtin.facultyFaculty of Science and Engineering
curtin.contributor.orcidMoffat, Caroline [0000-0002-6360-1933]
curtin.contributor.researcheridMoffat, Caroline [I-1058-2014]
curtin.identifier.article-numberARTN 182
dcterms.source.eissn1664-302X
curtin.contributor.scopusauthoridMoffat, Caroline [25936541300]


Files in this item

Thumbnail

This item appears in the following Collection(s)

Show simple item record

http://creativecommons.org/licenses/by/4.0/
Except where otherwise noted, this item's license is described as http://creativecommons.org/licenses/by/4.0/