Tagged mutations at the Tox1 locus of Cochliobolus heterostrophus by restriction enzyme-mediated integration.

Lu, Shunwen; Lyngholm, Linda K.; Yang, Ge; Bronson, C. R.; Yoder, O. C.; Turgeon, B. Gillian

doi:10.1073/pnas.91.26.12649

Cited by 224 publications

(135 citation statements)

References 22 publications

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“…The amplified DNA fragment was cloned into pGEM-T easy vector, creating pSTSF1128. A 2.1 kb fragment harbouring the hygromycin phosphotransferase B gene (HYG) cassette under the control of the Aspergillus nidulans trpC gene promoter and terminator was released from pUCATPH (Lu et al, 1994) by digestion with XbaI, blunt-ended with DNA polymerase I (New England BioLabs), and cloned into the NruI site of pSTSF1128, creating the disruption construct pTSF1128 (see Fig. 3).…”

Section: Methodsmentioning

confidence: 99%

Determination of a transcriptional regulator-like gene involved in biosynthesis of elsinochrome phytotoxin by the citrus scab fungus, Elsinoë fawcettii

Chung

Liao

2008

Microbiology

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Elsinochromes are nonhost-selective, light-activated, polyketide-derived toxins produced by many phytopathogenic Elsinoë species. We recently showed that the polyketide synthase-encoding gene EfPKS1 is essential for elsinochrome biosynthesis in the citrus scab fungus Elsinoë fawcettii. Sequence analysis beyond the EfPKS1 gene identified nine putative ORFs: four genes, designated RDT1, TSF1, PRF1 and ECT1, all encode polypeptides likely to have biosynthetic or efflux functions; five additional genes, OXR1 and EfHP1 to EfHP4, encode hypothetical proteins of unknown function. Northern-blot analysis revealed that expression of these genes in E. fawcettii was not completely correlated with accumulation of elsinochromes under nitrogen limitation, alkaline pH or high concentrations of glucose. Targeted disruption of the TSF1 gene, encoding a putative transcriptional activator, yielded fungal mutants unable to produce elsinochromes, and defective in both conidiation and expression of RDT1, EfPKS1, PRF1 and EfHP1, whereas expression of RDT1, TSF1, PRF1 and ECT1 was nearly abolished in EfPKS1-disrupted mutants. By contrast, expression of OXR1, EfHP2 and EfHP3 was not affected by disrupting either EfPKS1 or TSF1. Taken together, the results indicate that in addition to polyketide synthase, the products of TSF1 and other adjacent genes may also play a crucial role in elsinochrome production.

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Section: Methodsmentioning

confidence: 99%

Determination of a transcriptional regulator-like gene involved in biosynthesis of elsinochrome phytotoxin by the citrus scab fungus, Elsinoë fawcettii

Chung

Liao

2008

Microbiology

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“…1992; Lu et al 1994;Inoue et al 2001). This pathogen A genomic cosmid library of Mel02010 was constructed with causes vascular wilt of melon (Leach and Currence a cosmid vector pMLF2 using half-site fill-in reactions (Sambrook et al 1989).…”

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confidence: 99%

REN1Is Required for Development of Microconidia and Macroconidia, but Not of Chlamydospores, in the Plant Pathogenic FungusFusarium oxysporum

et al. 2004

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The filamentous fungus Fusarium oxysporum is a soil-borne facultative parasite that causes economically important losses in a wide variety of crops. F. oxysporum exhibits filamentous growth on agar media and undergoes asexual development producing three kinds of spores: microconidia, macroconidia, and chlamydospores. Ellipsoidal microconidia and falcate macroconidia are formed from phialides by basipetal division; globose chlamydospores with thick walls are formed acrogenously from hyphae or by the modification of hyphal cells. Here we describe rensa, a conidiation mutant of F. oxysporum, obtained by restrictionenzyme-mediated integration mutagenesis. Molecular analysis of rensa identified the affected gene, REN1, which encodes a protein with similarity to MedA of Aspergillus nidulans and Acr1 of Magnaporthe grisea. MedA and Acr1 are presumed transcription regulators involved in conidiogenesis in these fungi. The rensa mutant and REN1-targeted strains lack normal conidiophores and phialides and form rod-shaped, conidium-like cells directly from hyphae by acropetal division. These mutants, however, exhibit normal vegetative growth and chlamydospore formation. Nuclear localization of Ren1 was verified using strains expressing the Ren1-green fluorescent protein fusions. These data strongly suggest that REN1 encodes a transcription regulator required for the correct differentiation of conidiogenesis cells for development of microconidia and macroconidia in F. oxysporum.

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“…Briosi & Cavara. (Redman & Rodriguez, 1994), Cochliobolus heterostrophus (Drechsler) Drechsler (Lu et al, 1994), Aspergillus oryzae (Ahlb.) E. Cohn.…”

Section: Fig 2 -Sensibilidade Dementioning

confidence: 99%

Integração de pAN7-1 no genoma de Magnaporthe grisea mediada por enzima de restrição

Marchi¹,

Brommonschenkel²,

Queiroz

et al. 2006

Fitopatol. bras.

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Visando explorar a mutagênese insercional em Magnaporthe grisea, foram avaliadas a transformação dos protoplastos obtidos após adequação do protocolo e a eficiência da integração de pAN7-1 no genoma do ascomiceto na presença da enzima de restrição Hind III. Os protoplastos de M. grisea I-22 foram prontamente transformados para a resistência à higromicina. Quando o vetor linearizado com Hind III foi usado para transformar o fungo na presença de Hind III, a eficiência de transformação foi 1,1 a 8,1 vezes superior ao tratamento sem a adição da enzima. No geral, a melhor concentração de Hind III foi 5 unidades/reação de transformação. Tal concentração promoveu a produção média de 332 transformantes/µg de pAN7-1/10(7) protoplastos. A presença do gene de seleção hph no genoma de 18 indivíduos resistentes à higromicina foi confirmada por PCR.

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Tagged mutations at the Tox1 locus of Cochliobolus heterostrophus by restriction enzyme-mediated integration.

Cited by 224 publications

References 22 publications

Determination of a transcriptional regulator-like gene involved in biosynthesis of elsinochrome phytotoxin by the citrus scab fungus, Elsinoë fawcettii

Determination of a transcriptional regulator-like gene involved in biosynthesis of elsinochrome phytotoxin by the citrus scab fungus, Elsinoë fawcettii

REN1Is Required for Development of Microconidia and Macroconidia, but Not of Chlamydospores, in the Plant Pathogenic FungusFusarium oxysporum

Integração de pAN7-1 no genoma de Magnaporthe grisea mediada por enzima de restrição

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