Chemical and biochemical reactions of solanapyrone A, a toxin from the chickpea pathogen, Ascochyta rabiei (Pass.) Labr.

Bahti, Pazilat; Strange, Richard

doi:10.1016/j.pmpp.2004.05.002

Cited by 6 publications

(2 citation statements)

References 12 publications

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“…This work presents the first report of the successful genetic transformation of a fungus from the genus Ascochyta using Agrobacterium. Alternatively, by analogy with the detoxification of eutypine, an aldehydic toxin synthesised by Eutypa lata and affecting grapevine (Vitis vinifera), it may be possible to transform plants with an enzyme that reduces the aldehydic solanapyrone A to the less toxic solanapyrone B (Colrat et al, 1999a,b;Bahti & Strange, 2004). Future work with A. rabiei aims to extend the range of single-copy T-DNA tagged solanapyrone mutants by modification of the ATMT protocols (Mullins et al, 2001;Rho et al, 2001), recover the T-DNA insertions and characterize the mutagenized genes.…”

Section: Discussionmentioning

confidence: 99%

“…This may involve selection of plants with a well-developed glutathione/glutathione-Stransferase system for detoxification (Hamid & Strange, 2000). Alternatively, by analogy with the detoxification of eutypine, an aldehydic toxin synthesised by Eutypa lata and affecting grapevine (Vitis vinifera), it may be possible to transform plants with an enzyme that reduces the aldehydic solanapyrone A to the less toxic solanapyrone B (Colrat et al, 1999a,b;Bahti & Strange, 2004).…”

Section: Discussionmentioning

confidence: 99%

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Reduction in solanapyrone phytotoxin production byAscochyta rabieitransformed withAgrobacterium tumefaciens

Mogensen

Challen

Strange

2006

FEMS Microbiology Letters

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Agrobacterium tumefaciens was used to transform Ascochyta rabiei, the causal agent of chickpea blight. Employing a T-DNA containing a hygromycin resistance gene (hph), 908 transformants were obtained from germinated pycnidiospores on a selective medium containing hygromycin. Transformants were confirmed using PCR and Southern analyses and of four of these that were tested, two had integrated multicopies of the hph gene, one had two copies and one had a single insertion. Transformants were tested for the production of solanapyrone A toxin using a microtitre plate assay. Loss of toxin production by transformants was confirmed by reversed phase high-performance liquid chromatography. Sixteen transformants out of 668 tested produced significantly less solanapyrone A than the wild-type strain.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Reduction in solanapyrone phytotoxin production byAscochyta rabieitransformed withAgrobacterium tumefaciens

Mogensen

Challen

Strange

2006

FEMS Microbiology Letters

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Genetic transformation of Ascochyta rabiei using Agrobacterium-mediated transformation

White

Chen

2005

Curr Genet

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In order to study pathogenic mechanisms of the plant pathogen Ascochyta rabiei, conditions for efficient transformation using Agrobacterium-mediated transformation were investigated. Hygromycin B resistance (hph) was superior to geneticin resistance (nptII) for selecting transformants, and the hph gene was more efficiently expressed by the Aspergillus nidulans trpC promoter than by the Cauliflower mosaic virus 35S promoter CaMV35S. Co-cultivation on solid media for 72 h was optimal for generating transformants, but increasing the ratio of bacterial cells to conidia did not affect transformation efficiency. All hygromycin B-resistant transformants carried transfer-DNA (T-DNA) as determined by polymerase chain reaction (PCR) and the T-DNA integrations appeared to be random and in single copy as detected by Southern hybridization. Transformants remained resistant to hygromycin B in the absence of selection. Variations in colony morphology were observed in the presence of hygromycin B under different culture conditions, and a variety of altered phenotypes including reduced virulence were observed among 550 transformants. Inverse PCR was more efficient than TAIL-PCR in identifying flanking genomic sequences from T-DNA borders, and the possible causes are discussed. This transformation technique and recovery of flanking DNA using inverse PCR will provide a useful tool for genetic studies of A. rabiei.

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Functional Analyses of the Diels-Alderase Gene sol5 of Ascochyta rabiei and Alternaria solani Indicate that the Solanapyrone Phytotoxins Are Not Required for Pathogenicity

Kim

Park²,

Park

et al. 2015

MPMI

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Ascochyta rabiei and Alternaria solani, the causal agents of Ascochyta blight of chickpea (Cicer arietinum) and early blight of potato (Solanum tuberosum), respectively, produce a set of phytotoxic compounds including solanapyrones A, B, and C. Although both the phytotoxicity of solanapyrones and their universal production among field isolates have been documented, the role of solanapyrones in pathogenicity is not well understood. Here, we report the functional characterization of the sol5 gene, which encodes a Diels-Alderase that catalyzes the final step of solanapyrone biosynthesis. Deletion of sol5 in both Ascochyta rabiei and Alternaria solani completely prevented production of solanapyrones and led to accumulation of the immediate precursor compound, prosolanapyrone II-diol, which is not toxic to plants. Deletion of sol5 did not negatively affect growth rate or spore production in vitro, and led to overexpression of the other solanapyrone biosynthesis genes, suggesting a possible feedback regulation mechanism. Phytotoxicity tests showed that solanapyrone A is highly toxic to several legume species and Arabidopsis thaliana. Despite the apparent phytotoxicity of solanapyrone A, pathogenicity tests showed that solanapyrone-minus mutants of Ascochyta rabiei and Alternaria solani were equally virulent as their corresponding wild-type progenitors, suggesting that solanapyrones are not required for pathogenicity.

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Chemical and biochemical reactions of solanapyrone A, a toxin from the chickpea pathogen, Ascochyta rabiei (Pass.) Labr.

Cited by 6 publications

References 12 publications

Reduction in solanapyrone phytotoxin production byAscochyta rabieitransformed withAgrobacterium tumefaciens

Reduction in solanapyrone phytotoxin production byAscochyta rabieitransformed withAgrobacterium tumefaciens

Genetic transformation of Ascochyta rabiei using Agrobacterium-mediated transformation

Functional Analyses of the Diels-Alderase Gene sol5 of Ascochyta rabiei and Alternaria solani Indicate that the Solanapyrone Phytotoxins Are Not Required for Pathogenicity

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