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
Pao Theen SeeCaroline S Moffat

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 pr...Continue Reading

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Citations

Jan 10, 2020·TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik·Beatrice CorsiJames Cockram
May 18, 2020·Brazilian Journal of Microbiology : [publication of the Brazilian Society for Microbiology]·Na ZhanAnshan Shan
Jan 11, 2020·Frontiers in Plant Science·Sana KamelReem Aboukhaddour
Mar 18, 2020·Frontiers in Plant Science·Bohan WeiReem Aboukhaddour
Apr 14, 2019·Planta·Pramod PrasadSubodh Kumar
Jun 2, 2021·TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik·Eric G DinglasanLee T Hickey

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Datasets Mentioned

BETA
PZD27634

Methods Mentioned

BETA
PCR
NMR
affinity purification

Software Mentioned

EGA Kidman

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