A Catalytic Domain of a Cyclic Peptide Synthetase that is Specific for the Apple Pathotype of Alternaria Alternata and its Possible Involvement in Host-Specific AM-Toxin Production

1998 ◽  
pp. 53-61
Author(s):  
Y. Itoh ◽  
R. Kiyohara ◽  
Y. Kawamoto ◽  
M. Kodama ◽  
H. Otani ◽  
...  
Keyword(s):  
Alternaria Alternata ◽  
Catalytic Domain ◽  
Cyclic Peptide ◽  
Toxin Production ◽  
Peptide Synthetase

scholarly journals Cloning and Characterization of a Cyclic Peptide Synthetase Gene from Alternaria alternata Apple Pathotype Whose Product Is Involved in AM-Toxin Synthesis and Pathogenicity

2000 ◽  
Vol 13 (7) ◽  
pp. 742-753 ◽  
Author(s):  
R. D. Johnson ◽  
L. Johnson ◽  
Y. Itoh ◽  
M. Kodama ◽  
H. Otani ◽  
...  
Keyword(s):  
Alternaria Alternata ◽  
Cyclic Peptide ◽  
Genomic Library ◽  
Disease Symptoms ◽  
Peptide Synthetases ◽  
Peptide Synthetase ◽  
Primary Determinant ◽  
Apple Cultivars ◽  
Alternaria Blotch

Alternaria alternata apple pathotype causes Alternaria blotch of susceptible apple cultivars through the production of a cyclic peptide host-specific toxin, AM-toxin. PCR (polymerase chain reaction), with primers designed to conserved domains of peptide synthetase genes, amplified several products from A. alternata apple pathotype that showed high similarity to other fungal peptide synthetases and were specific to the apple pathotype. Screening of a Lambda Zap genomic library with these PCR-generated probes identified overlapping clones containing a complete cyclic peptide synthetase gene of 13.1 kb in length with no introns. Disruption of this gene, designated AM-toxin synthetase (AMT), by transformation of wild-type A. alternata apple pathotype with disruption vectors resulted in toxin-minus mutants, which were also unable to cause disease symptoms on susceptible apple cultivars. AM-toxin synthetase is therefore a primary determinant of virulence and specificity in the A. alternata apple pathotype/apple interaction.

scholarly journals A Fatty Acid Synthase Gene in Cochliobolus carbonum Required for Production of HC-Toxin, Cyclo(d-Prolyl-l-Alanyl- d-Alanyl- l-2-Amino-9,10-Epoxi-8-Oxodecanoyl)

1997 ◽  
Vol 10 (2) ◽  
pp. 207-214 ◽  
Author(s):  
Joong-Hoon Ahn ◽  
Jonathan D. Walton
Keyword(s):  
Fatty Acid ◽  
Fatty Acid Synthase ◽  
Cyclic Peptide ◽  
Decanoic Acid ◽  
Toxin Production ◽  
Gene Encoding ◽  
Cochliobolus Carbonum ◽  
Primary Amino ◽  
Hc Toxin

The fungal maize pathogen Cochliobolus carbonum produces a phytotoxic and cytostatic cyclic peptide, HC-toxin, of structure cyclo(D-prolyl-L-alanyl-D-alanyl-L-Aeo), in which Aeo stands for 2-amino-9,10-epoxi-8-oxodecanoic acid. Here we report the isolation of a gene, TOXC, that is present only in HC-toxin-producing (Tox2+) fungal strains. TOXC is present in most Tox2+ strains in three functional copies, all of which are on the same chromosome as the gene encoding HC-toxin synthetase. When all copies of TOXC are mutated by targeted gene disruption, the fungus grows and sporulates normally in vitro but no longer makes HC-toxin and is not pathogenic, indicating that TOXC has a specific role in HC-toxin production and hence virulence. The TOXC mRNA is 6.5 kb and the predicted product has 2,080 amino acids and a molecular weight of 233,000. The primary amino acid sequence is highly similar (45 to 47% identity) to the β subunit of fatty acid synthase from several lower eukaryotes, and contains, in the same order as in other β subunits, domains predicted to encode acetyl transferase, enoyl reductase, dehydratase, and malonyl-palmityl transferase. The most plausible function of TOXC is to contribute to the synthesis of the decanoic acid backbone of Aeo.

scholarly journals Studies on variability in Alternaria alternata (Kessler) causing leaf blight of Isabgol (Plantago ovata)

2015 ◽  
Vol 12 (2) ◽  
pp. 63-70 ◽  
Author(s):  
RK Meena ◽  
SS Sharma ◽  
S Singh
Keyword(s):  
Alternaria Alternata ◽  
Mycelial Growth ◽  
Toxin Production ◽  
Plantago Ovata ◽  
Mycelium Growth ◽  
Highly Pathogenic ◽  
Fast Growing ◽  
Conidial Morphology ◽  
Incubation Periods ◽  
Pathogenic Variability

All the five isolates of Alternaria alternata isolated from different agro climate zone of Rajasthan were tested for their variability in terms of cultural, conidial, pathogenic characteristics and toxin production. All the five isolates differed in cultural characters i.e. dark black colored and very fast mycelial growth with smooth margins (90.00 mm), light black with white at centre and fast growing (80.00 mm), dark brown and medium mycelium growth with smooth margins (75.00 mm), black colored, medium flat mycelial growth with smooth margins (68.00 mm) and white with slightly black in colour with slow mycelial growth (65.00 mm) were observed in Aa-1, Aa-2, Aa-3, Aa-4 and Aa-5 respectively. The variability in conidial morphology of five different isolates was simple, septate, pale to dark brown in colour, often geniculate with one conidial scar. In respect of pathogenic variability, showed significant variations in terms of disease intensity and incubation periods. The isolates Aa-1 was highly pathogenic on Isabgol cv. RI-89 under artificial inoculation conditions showing 52.12% disease intensity followed by Aa- 3 ,Aa-2, Aa-4 and Aa-5 isolates. The variability in toxin production was reflected in terms of time taken in inducing wilting symptoms of Isabgol cuttings. Isolate Aa-1 was highly toxic followed by isolates Aa-2, Aa-3, Aa-4 and Aa-5. DOI: http://dx.doi.org/10.3329/sja.v12i2.21918 SAARC J. Agri., 12(2): 63-70 (2014)

scholarly journals Distribution and Characterization of AKT Homologs in the Tangerine Pathotype of Alternaria alternata

2000 ◽  
Vol 90 (7) ◽  
pp. 762-768 ◽  
Author(s):  
A. Masunaka ◽  
A. Tanaka ◽  
T. Tsuge ◽  
T. L. Peever ◽  
L. W. Timmer ◽  
...  
Keyword(s):  
Alternaria Alternata ◽  
Sequence Similarity ◽  
Structural Similarity ◽  
Toxin Production ◽  
Brown Spot ◽  
Japanese Pear ◽  
Black Rot ◽  
Acid Moiety ◽  
High Sequence Similarity ◽  
Rough Lemon

The tangerine pathotype of Alternaria alternata produces a host-selective toxin (HST), known as ACT-toxin, and causes Alternaria brown spot disease of citrus. The structure of ACT-toxin is closely related to AK- and AF-toxins, which are HSTs produced by the Japanese pear and strawberry pathotypes of A. alternata, respectively. AC-, AK-, and AF-toxins are chemically similar and share a 9,10-epoxy-8-hydroxy-9-methyl-decatrienoic acid moiety. Two genes controlling AK-toxin biosynthesis (AKT1 and AKT2) were recently cloned from the Japanese pear pathotype of A. alternata. Portions of these genes were used as heterologous probes in Southern blots, that detected homologs in 13 isolates of A. alternata tangerine pathotype from Minneola tangelo in Florida. Partial sequencing of the homologs in one of these isolates demonstrated high sequence similarity to AKT1 (89.8%) and to AKT2 (90.7%). AKT homologs were not detected in nine isolates of A. alternata from rough lemon, six isolates of nonpathogenic A. alternata, and one isolate of A. citri that causes citrus black rot. The presence of homologs in the Minneola isolates and not in the rough lemon isolates, nonpathogens or black rot isolates, correlates perfectly to pathogenicity on Iyo tangerine and ACT-toxin production. Functionality of the homologs was demonstrated by detection of transcripts using reverse transcription-polymerase chain reaction (RT-PCR) in total RNA of the tangerine pathotype of A. alternata. The high sequence similarity of AKT and AKT homologs in the tangerine patho-type, combined with the structural similarity of AK-toxin and ACT-toxin, may indicate that these homologs are involved in the biosynthesis of the decatrienoic acid moiety of ACT-toxin.

scholarly journals Characterization of the Ustilago maydis sid2 Gene, Encoding a Multidomain Peptide Synthetase in the Ferrichrome Biosynthetic Gene Cluster

2001 ◽  
Vol 183 (13) ◽  
pp. 4040-4051 ◽  
Author(s):  
Walter M. Yuan ◽  
Guillaume D. Gentil ◽  
Allen D. Budde ◽  
Sally A. Leong
Keyword(s):  
Amino Acids ◽  
Cyclic Peptide ◽  
Ustilago Maydis ◽  
Nonribosomal Peptide Synthetase ◽  
Hybridization Analysis ◽  
Northern Hybridization ◽  
Nucleotide Sequencing ◽  
Biosynthetic Gene ◽  
Nonribosomal Peptide ◽  
Peptide Synthetase

ABSTRACT Ustilago maydis, the causal agent of corn smut disease, acquires and transports ferric ion by producing the extracellular, cyclic peptide, hydroxamate siderophores ferrichrome and ferrichrome A. Ferrichrome biosynthesis likely proceeds by hydroxylation and acetylation of l-ornithine, and later steps likely involve covalently bound thioester intermediates on a multimodular, nonribosomal peptide synthetase. sid1 encodesl-ornithine N 5-oxygenase, which catalyzes hydroxylation of l-ornithine, the first committed step of ferrichrome and ferrichrome A biosynthesis in U. maydis. In this report we characterize sid2, another biosynthetic gene in the pathway, by gene complementation, gene replacement, DNA sequence, and Northern hybridization analysis. Nucleotide sequencing has revealed that sid2 is located 3.7 kb upstream of sid1 and encodes an intronless polypeptide of 3,947 amino acids with three iterated modules of an approximate length of 1,000 amino acids each. Multiple motifs characteristic of the nonribosomal peptide synthetase protein family were identified in each module. A corresponding iron-regulated sid2 transcript of 11 kb was detected by Northern hybridization analysis. By contrast, constitutive accumulation of this large transcript was observed in a mutant carrying a disruption of urbs1, a zinc finger, GATA family transcription factor previously shown to regulate siderophore biosynthesis in Ustilago. Multiple GATA motifs are present in the intergenic region between sid1 and sid2, suggesting bidirectional transcription regulation by urbs1of this pathway. Indeed, mutation of two of these motifs, known to be important to regulation of sid1, altered the differential regulation of sid2 by iron.

scholarly journals Role of the Host-Selective ACT-Toxin Synthesis Gene ACTTS2 Encoding an Enoyl-Reductase in Pathogenicity of the Tangerine Pathotype of Alternaria alternata

2010 ◽  
Vol 100 (2) ◽  
pp. 120-126 ◽  
Author(s):  
Naoya Ajiro ◽  
Yoko Miyamoto ◽  
Akira Masunaka ◽  
Takashi Tsuge ◽  
Mikihiro Yamamoto ◽  
...  
Keyword(s):  
Alternaria Alternata ◽  
Essential Gene ◽  
Toxin Production ◽  
Brown Spot ◽  
Gene Encoding ◽  
Bac Clone ◽  
Biosynthesis Gene Cluster ◽  
Enoyl Reductase ◽  
Alternaria Brown Spot ◽  
Rapid Amplification

The tangerine pathotype of Alternaria alternata produces host-selective ACT-toxin and causes Alternaria brown spot disease of tangerines and tangerine hybrids. Sequence analysis of a genomic BAC clone identified a previously uncharacterized portion of the ACT-toxin biosynthesis gene cluster (ACTT). A 1,034-bp gene encoding a putative enoyl-reductase was identified by using rapid amplification of cDNA ends and polymerase chain reaction and designated ACTTS2. Genomic Southern blots demonstrated that ACTTS2 is present only in ACT-toxin producers and is carried on a 1.9 Mb conditionally dispensable chromosome by the tangerine pathotype. Targeted gene disruption of ACTTS2 led to a reduction in ACT-toxin production and pathogenicity, and transcriptional knockdown of ACTTS2 using RNA silencing resulted in complete loss of ACT-toxin production and pathogenicity. These results indicate that ACTTS2 is an essential gene for ACT-toxin biosynthesis in the tangerine pathotype of A. alternata and is required for pathogenicity of this fungus.

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