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Cassava brown streak virus Ham1 protein hydrolyses mutagenic nucleotides and is a necrosis determinant.

ABSTRACT: Cassava brown streak disease (CBSD) is a leading cause of cassava losses in East and Central Africa, and is currently having a severe impact on food security. The disease is caused by two viruses within the Potyviridae family: Cassava brown streak virus (CBSV) and Ugandan cassava brown streak virus (UCBSV), which both encode atypical Ham1 proteins with highly conserved inosine triphosphate (ITP) pyrophosphohydrolase (ITPase) domains. ITPase proteins are widely encoded by plant, animal, and archaea. They selectively hydrolyse mutagenic nucleotide triphosphates to prevent their incorporation into nucleic acid and thereby function to reduce mutation rates. It has previously been hypothesized that U/CBSVs encode Ham1 proteins with ITPase activity to reduce viral mutation rates during infection. In this study, we investigate the potential roles of U/CBSV Ham1 proteins. We show that both CBSV and UCBSV Ham1 proteins have ITPase activities through in vitro enzyme assays. Deep-sequencing experiments found no evidence of the U/CBSV Ham1 proteins providing mutagenic protection during infections of Nicotiana hosts. Manipulations of the CBSV_Tanza infectious clone were performed, including a Ham1 deletion, ITPase point mutations, and UCBSV Ham1 chimera. Unlike severely necrotic wild-type CBSV_Tanza infections, infections of Nicotiana benthamiana with the manipulated CBSV infectious clones do not develop necrosis, indicating that that the CBSV Ham1 is a necrosis determinant. We propose that the presence of U/CBSV Ham1 proteins with highly conserved ITPase motifs indicates that they serve highly selectable functions during infections of cassava and may represent a euphorbia host adaptation that could be targeted in antiviral strategies.

SUBMITTER: Tomlinson KR

PROVIDER: S-EPMC6640186 | biostudies-literature | 2019 Aug

REPOSITORIES: biostudies-literature

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Cassava brown streak virus Ham1 protein hydrolyses mutagenic nucleotides and is a necrosis determinant.

Tomlinson Katie R KR Pablo-Rodriguez José Luis JL Bunawan Hamidun H Nanyiti Sarah S Green Patrick P Miller Josie J Alicai Titus T Seal Susan E SE Bailey Andy M AM Foster Gary D GD

Molecular plant pathology 20190601 8

Cassava brown streak disease (CBSD) is a leading cause of cassava losses in East and Central Africa, and is currently having a severe impact on food security. The disease is caused by two viruses within the Potyviridae family: Cassava brown streak virus (CBSV) and Ugandan cassava brown streak virus (UCBSV), which both encode atypical Ham1 proteins with highly conserved inosine triphosphate (ITP) pyrophosphohydrolase (ITPase) domains. ITPase proteins are widely encoded by plant, animal, and archa ...[more]

PMID: 31154674

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Project description:Cassava brown streak disease (CBSD) caused by the rapidly evolving cassava brown streak viruses (CBSVs), causes immense yield losses to the cassava value chain in eastern and southern Africa. Western Africa, another region that heavily depends on cassava is under eminent threat from CBSD. Resistance breeding is the best practical solution. However, complexities associated with CBSD resistance screening i.e., variable root sampling units, limit systematic attainment of genetic progress. Accordingly, we compared efficiency of five CBSD root necrosis assessment methods to guide selection: cassava brown streak disease root incidence (CBSDRi), cassava brown streak disease root severity (CBSDRs), cassava brown streak disease root severity computed as harmonic mean (CBSD-Harmonic), proportion-based root necrosis index (CBSD-proportion), and standardized root necrosis index (CBSD-standardized). The indexes (CBSD-proportion and CBSD-standardized) correct for variable sample size. We analyzed CBSD root necrosis data of 256 clones evaluated across 12 environments. Higher and variable standard errors were associated with root severity score 1 (no CBSD root necrosis). Lowest and highest plot-based heritability were respectively registered for CBSD-standardized (0.22) and CBSD-proportion (0.71). CBSDRs was only positively correlated with CBSDRi (r = 0.92) and CBSD-Harmonic (r = 0.97). Using best linear unbiased predictions (BLUPs), we ranked the top 15 CBSD resistant clones; only one clone (UG130014) featured in all the five assessment methods; two clones (UG130006 and UG120156) featured in four (CBSD-Harmonic, CBSDRi, CBSDRs, and CBSD-standardized); and five clones (UG120180, UG120063, UG130002, UG130033, and UG120183) featured in three methods (CBSD-Harmonic, CBSDRi, and CBSDRs). Influence of sample size was also quantified by sub-setting and analyzing CBSDRs data to have plots with at least 40 or 30 roots. Data stabilization was evident in plots with 30 roots. The significant influence of root sample sizes on overall ranking of clones, justifies the use of CBSD root necrosis indexes in early selection stages i.e., seedling and/or clonal trials, that are often characterized by high variations in roots assessed per plot. It is expected that this information will provide a foundation for harmonizing and/or optimizing on-going and future CBSD resistance breeding efforts.

Dataset Information

Cassava brown streak virus Ham1 protein hydrolyses mutagenic nucleotides and is a necrosis determinant.

Publications

Cassava brown streak virus Ham1 protein hydrolyses mutagenic nucleotides and is a necrosis determinant.

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