Previous Article | Next Article 
Applied and Environmental Microbiology, July 2001, p. 2966-2972, Vol. 67, No. 7
School of Agricultural Biotechnology, Seoul
National University, Suwon 441-744,1 and
Division of Life Sciences, Soonchunhyang University, Asan
336-745,2 Korea
Received 22 January 2001/Accepted 12 April 2001
Gibberella zeae, a major cause of cereal scab, may be
divided into two chemotypes based on production of the trichothecenes deoxynivalenol (DON) and nivalenol (NIV). We cloned and sequenced the
gene cluster for trichothecene biosynthesis from each chemotype. G. zeae H-11 is a DON producer isolated from corn, and
G. zeae 88-1 is a NIV producer from barley. We sequenced a
23-kb gene cluster from H-11 and a 26-kb cluster from 88-1, along with
the unlinked Tri101 genes. Each gene cluster contained 10 Tri gene homologues in the same order and transcriptional
directions as those of Fusarium sporotrichioides. Between
H-11 and 88-1 all of the Tri homologues except
Tri7 were conserved, with identities ranging from 88 to
98% and 82 to 99% at the nucleotide and amino acid levels,
respectively. The Tri7 sequences were only 80% identical at the nucleotide level. We aligned the Tri7 genes and
found that the Tri7 open reading frame of H-11 carried
several mutations and an insertion containing 10 copies of an 11-bp
tandem repeat. The Tri7 gene from 88-1 carried neither the
repeat nor the mutations. We assayed 100 G. zeae isolates
of both chemotypes by PCR amplification with a primer pair derived from
the Tri7 gene and could differentiate the chemotypes by
polyacrylamide gel electrophoresis. The PCR-based method developed in
this study should provide a simple and reliable diagnostic tool for
differentiating the two chemotypes of G. zeae.
Gibberella zeae
(Schwein.) Petch (anamorph: Fusarium graminearum Schwabe) is
an important pathogen of cereal crops in many areas of the world. The
fungus causes head and seedling blight of small grains, such as wheat
and barley; ear and stalk rot of corn; and stem rot of carnation
(7, 20, 28). Head blight and ear rot reduce the yield of
grain, and harvested grain is often contaminated with mycotoxins, such
as trichothecenes and zearalenone (23). The fungus
produces 8-ketotrichothecenes, including deoxynivalenol
(DON), 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, nivalenol
(NIV), and 4-acetylnivalenol (4-ANIV), as well as an estrogenic
mycotoxin, zearalenone (27, 35). Of these
8-ketotrichothecenes, DON and NIV are frequently found in cereals
harvested in Korea and Japan (17, 38, 40). Compared to
DON, NIV is present at higher levels in cereals and exhibits greater
toxicity (33). Therefore, NIV is of greater concern in
these countries.
Based on 8-ketotrichothecene production, Ichinoe et al.
(15) divided G. zeae into two chemotypes
groups. The DON chemotype produces DON and acetyl-DON, and the NIV
chemotype produces NIV and 4-ANIV. These chemotypes appear to differ in
geographic distribution. The NIV chemotype has been reported in several
countries of Africa, Asia, and Europe (10, 14, 22, 35,
37), but it has not been reported in North America (1,
27). Since the NIV chemotype is of great concern in the
countries where it has appeared, an efficient system for
differentiating DON and NIV chemotypes is desired. Recently
O'Donnell et al. (30) divided global population of
G. zeae into seven biogeographically structured lineages
based on phylogenetic analysis using six gene genealogies. The two
chemotypes, however, do not appear to correlate with the global
phylogenetic structure of G. zeae.
Although the geographic distribution of DON and NIV chemotypes has been
well studied, little is known about the genetic basis of
8-ketotrichothecene production by these chemotypes. However, the molecular genetics of the production of T-2 toxin by Fusarium sporotrichioides has been studied intensively, using various
mutant strains blocked at specific steps in the
trichothecene pathway (9, 13). Many
trichothecene biosynthesis genes are localized in a gene
cluster comprising at least 10 genes. These genes include those
encoding trichodiene synthetase (Tri5)
(11), P450 oxygenases (Tri4 and
Tri11) (2, 12), acetyltransferase
(Tri3) (25), a transcription factor
(Tri6) (32), a toxin efflux pump
(Tri12) (3), and several unidentified
hypothetical proteins (Tri7, Tri8,
Tri9, and Tri10) (13, 26). Another
acetyltransferase (Tri101) (19) is unlinked to
the cluster. Homologues of Tri5, Tri6,
Tri11, Tri12, and Tri101 have been
previously found in G. zeae (18, 24, 31, 39),
and all of the genes exhibited high degrees of sequence homology to
those of F. sporotrichioides. A homologue of the F. sporotrichioides Tri12 was designated Tri102 in
G. zeae (39).
In this study we built on the F. sporotrichioides results to
analyze the trichothecene biosynthesis gene clusters in the DON and NIV chemotypes of G. zeae. The objectives of the study
were to identify genetic differences between the trichothecene
biosynthetic pathways of the two chemotypes, and to develop a rapid
method for G. zeae chemotype identification based on PCR analysis.
Strains, media, and culture conditions.
G. zeae
strains H-11 (a DON producer) and 88-1 (a NIV producer) were isolated
from Korean corn and barley, respectively, and used for sequence
analysis. For PCR assays, 100 field isolates of G. zeae were
used. Of these isolates, 25 DON producers were from the United States.
The remaining 25 DON and 50 NIV producers were from a previous study of
Korean strains (35). Fungi were cultured on potato
dextrose agar (Difco Laboratories, Detroit, Mich.) and preserved as
25% glycerol stock cultures frozen at DNA manipulations and PCR conditions.
Fungal genomic DNA was
prepared as previously described (16). E. coli
colonies carrying recombinant plasmids were screened by a single-tube
mini-prep method (21). For sequencing, plasmids were
purified from 5-ml E. coli cultures using a Qiagen kit
(Qiagen Inc., Valencia, Calif.). Standard procedures were used for
restriction endonuclease digestions, ligations, and agarose or
polyacrylamide gel electrophoresis (34).
0099-2240/01/$04.00+0 DOI: 10.1128/AEM.67.7.2966-2972.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Identification of Deoxynivalenol- and
Nivalenol-Producing Chemotypes of Gibberella zeae by
Using PCR
ABSTRACT
Top
Abstract
Introduction
Materials and Methods
Results
Discussion
References
INTRODUCTION
Top
Abstract
Introduction
Materials and Methods
Results
Discussion
References
MATERIALS AND METHODS
Top
Abstract
Introduction
Materials and Methods
Results
Discussion
References
80°C. To isolate genomic
DNA, fungal conidia were inoculated into 100 ml of complete liquid
medium (8) at 106 per ml. Cultures were
incubated in 250-ml Erlenmyer flasks at 25°C for 48 h on a
rotary shaker (200 rpm), after which mycelia were harvested and
lyophilized. Escherichia coli strains were grown on
Luria-Bertani agar or liquid medium supplemented with ampicillin (75 µg/ml).
20°C. For PCRs, about 50 ng of genomic DNA was
used as a template in a 50-µl reaction mixture containing Ex
Taq PCR buffer, which contains 2 mM MgCl2 (TaKaRa Biomedicals, Shiga, Japan), deoxynucleoside triphosphate mixture (0.2 mM each), a 2 µM concentration of each primer, and 1.25 U of Ex Taq (TaKaRa Biomedicals). PCRs were performed in a
thermal cycler (MJ Research, Inc., Waltham, Mass.) set to the following: denaturation at 94°C for 2 min; 30 cycles of 94°C for 30 s, 60°C for 1 min, and 72°C for 2 min 30 s; and a
final extension step at 72°C for 10 min. After PCR was completed, PCR
mixtures were stored at 4°C.
TABLE 1.
Primers used for PCR-based cloning
Amplification of Tri gene clusters.
We amplified
Tri genes from both strains (Fig.
1) by using several sets of degenerate
primers that were based on the known sequences of Tri genes
from G. zeae and F. sporotrichioides. Most of the
degenerate primer pairs successfully amplified fragments of the
expected sizes from the two strains. When necessary, primers were
redesigned according to the consensus sequences of already-acquired Tri genes (e.g., primer pairs 5-6 and 5'-6' in Table 1 and
Fig. 1). The PCR product sequences were used to design specific primers for amplifying regions flanking the amplified Tri genes. A
PCR was performed using two primers, each derived from different
Tri genes based on the assumption that the order of the
Tri genes in the G. zeae and F. sporotrichioides clusters is the same. For example, PCR using
primers 31 and 43 successfully amplified the region between
Tri6 and Tri5 from H-11 (Fig. 1). When this
strategy failed, inverse PCR using primers derived from the known
sequences was performed as previously described (29, 41).
These PCR strategies yielded various sizes of DNA fragments sufficient
for construction of contigs for both strains.
|
Cloning and sequencing. The amplified PCR products were analyzed by agarose gel electrophoresis. PCR products of the expected sizes were cloned into pCR2.1TOPO using the TOPO TA cloning kit (Invitrogen, San Diego, Calif.). Sequencing of the inserts in pCR2.1 TOPO was initiated with M13 reverse and forward primers and then extended using specific primers corresponding to the newly sequenced regions. DNA sequencing was done at the National Instrumentation Center for Environmental Management, Seoul National University, Suwon, Korea, using an ABI377 automated DNA sequencer (Applied Biosystems Inc., Foster City, Calif.). Primers for sequencing were designed using the PrimerSelect program (DNASTAR, Inc., Madison, Wis.). Sequences were assembled using the SeqMan program (DNASTAR, Inc.) and analyzed with the MegAlign and MapDraw programs (DNASTAR, Inc.). BLAST (4) searches were done against the NCBI and GenBank databases.
PCR assays. To amplify the inserted region of Tri7 from 50 DON- and 50 NIV-producing isolates, we designed a specific set of primers. The sequence of primer GzTri7/f1 (forward) is 5'-GGCTTTACGACTCCTCAACAATGG-3', and the sequence of primer GzTri7/r1 (reverse) is 5'-AGAGCCCTGCGAAAG(C/T)ACTGGTGC-3'. PCRs were performed in a thermal cycler set to the following: denaturation at 94°C for 2 min; 30 cycles of 94°C for 1 min, 60°C for 30 s, and 72°C for 1 min; and a final extension at 72°C for 10 min. After PCR was completed, PCR mixtures were stored at 4°C. PCR products were electrophoresed on 5% polyacrylamide gels; the products of the expected size (~160 bp) were purified using a QIAquick PCR purification kit (Qiagen Inc.) and directly sequenced using primers GzTri7/f1 and GzTri7/r1.
Nucleotide sequence accession numbers. The sequences of the trichothecene biosynthesis gene clusters we obtained from G. zeae 88-1 and H-11 have been deposited in GenBank under accession numbers AF336365 and AF336366, respectively.
| |
RESULTS |
|---|
|
Top
Abstract
Introduction
Materials and Methods
Results
Discussion
References
|
|---|
Structural organization of trichothecene biosynthesis gene clusters from DON and NIV chemotypes. Construction of contigs using the sequenced PCR fragments yielded 26-and 23-kb gene clusters from G. zeae 88-1 and H-11, respectively. Each cluster carried 8 open reading frames (ORFs), all of which were readily identified by sequence comparisons with current databases. Two additional ORFs (designated Tri9 and Tri10) were found between Tri5 and Tri11 as those in the trichothecene gene cluster of F. sporotrichioides (Fig. 1). The order and transcription directions of the ORFs are identical for the two G. zeae chemotypes and F. sporotrichioides.
Comparative sequence analysis.
We compared the sizes of the
ORFs, putative introns, and adjacent noncoding DNA regions (Table
2) and the percent identities of the ORFs
and their flanking sequences (Table 3) in
the two G. zeae Tri clusters and other strains of G. zeae and F. sporotrichioides. All of the Tri
genes except Tri7 exhibited significant conservation between
the two strains, ranging from 88% (Tri8) to 98%
(Tri101) identity at the nucleotide level and from 82%
(Tri8) to 99% (Tri101) at the amino acid level.
The flanking regions exhibited less nucleotide sequence identity than
the ORFs, ranging from 57% (region between Tri3 and
Tri4) to 89% (region between Tri5 and
Tri11). In particular, strain H-11 lacked a 226-bp stretch
in the noncoding region between Tri3 and Tri4
that was present in strain 88-1 (data not shown), giving this region
the lowest percent identity overall.
|
|
Alignment of Tri7 ORFs from G. zeae DON and NIV chemotypes and F. sporotrichioides. Unlike the other Tri ORFs, the Tri7 ORFs from the two G. zeae chemotypes and from F. sporotrichioides exhibited striking differences. First, the nucleotide sequence of the H-11 Tri7 ORF exhibited only 80% identity to that of 88-1 and 64% identity to that of F. sporotrichioides (Table 3). This level of conservation was significantly lower than that found for the other Tri ORFs. Second, alignment of the nucleotide sequences of these three Tri7 ORFs revealed several alterations present only in H-11 (data not shown). For example, a substitution in a potential start codon in H-11 appeared to have occurred, causing a deficient translation start signal (by comparison with F. sporotrichioides). The H-11 Tri7 nucleotide sequence also appeared to have had several deletions, an addition, or other substitutions, resulting in frame shifts that created an internal stop in the putative Tri7 amino acid sequence. Due to these features, comparisons of the H-11 Tri7 amino acid sequence with those of 88-1 and F. sporotrichioides could not be made. Finally, the most substantial differences between the H-11 Tri7 gene and the other two Tri7 genes were found within a putative intron sequence. The Tri7 ORFs of 88-1 and F. sporotrichioides were interrupted by a putative intron at the same position, identified from the consensus signals for intron splicing (5'-GTAAT~TAG-3'). The corresponding region of H-11 was, however, much longer (151 bp) than those of 88-1 and F. sporotrichioides (51 bp each). This size difference was due to the presence of 10 tandem repeats of an 11-bp nucleotide stretch (CACAATATTAG) within that region of the H-11 Tri7 sequence. These repeats were not present in either 88-1 or F. sporotrichioides.
Amplification of the inserted regions of Tri7 genes
from field isolates.
Selected regions spanning the Tri7
insertion from G. zeae field isolates were PCR amplified
using primers GzTri7/f1 and GzTri7/r1. Amplification yielded products
of various sizes from DON chemotypes but yielded products of only one
size (~160 bp) from all NIV chemotypes tested (Fig.
2). The amplified DON chemotype products
were of distinctly lower mobility than the NIV chemotype products on
5% polyacrylamide gels. Direct sequencing of the PCR products revealed that the actual sizes of the products from DON chemotypes varied, ranging from 173 to 327 bp, depending on the number of 11-bp repeats within each sequence, whereas the products from NIV chemotypes were
identical in size (161 bp) due to a lack of the repeat. The inserted
repeats in the Tri7 sequences varied from 2 to 16 copies for
the 50 DON-producing isolates (data not shown). This variation was not,
however, related to their geographical origins.
|
| |
DISCUSSION |
|---|
|
Top
Abstract
Introduction
Materials and Methods
Results
Discussion
References
|
|---|
We determined the sequence of the trichothecene biosynthesis gene cluster from DON and NIV-producing G. zeae isolates. We used these sequences to identify potential changes that might be responsible for differences in 8-ketotrichothecene biosynthesis.
Based on sequence differences, we think that the Tri7 gene may be one of the elements responsible for the difference in trichothecene production between the two chemotypes, as either no Tri7 protein or a truncated version is synthesized in H-11. These features are conserved in all of the DON isolates tested (data not shown), suggesting that the Tri7 gene is nonfunctional in all DON chemotypes. To confirm the role of Tri7 protein in G. zeae trichothecene biosynthesis, functional studies are needed. Previous experiments suggest that Tri7 protein is required for acetylation of the hydroxyl group at C-4 of T-2 toxin produced by F. sporotrichioides (13, 26). If the function of Tri7 in G. zeae is the same as in F. sporotrichioides, Tri7 protein is not required for DON production but is required for acetylation of the hydroxyl group at C-4 of NIV to convert to 4-ANIV; NIV chemotypes usually produce both NIV and 4-ANIV. Recently, an additional homologue of oxygenase (TriD) was found immediately upstream of Tri102 in both F. sporotrichioides and a DON-producing strain of G. zeae. However, sequence comparison showed that the TriD gene of the DON chemotype was defective (6). If TriD is functional in the NIV chemotypes as in F. sporotrichioides, both TriD and Tri7 would be determinants for biosynthesis of NIV and 4-ANIV in G. zeae.
The presence of different numbers of repeats within the Tri7 gene sequences of DON isolates suggests certain possibilities. The defective Tri7 gene in DON chemotypes of G. zeae may no longer be under selection pressure. This lack of selection would allow mutations to accumulate. Another possible mechanism for the variable number of repeats among the DON isolates is unequal crossing over during sexual recombination (5). To test this possibility, it is necessary to evaluate ascospore progeny from G. zeae DON chemotypes and see if the number of repeats present in the progeny is the same as in the parents. The Tri7 gene sequence could also be used as a marker to assess genetic diversity among DON isolates as well as to distinguish the chemotypes of G. zeae.
O'Donnell et al. (30) estimated global genetic diversity of G. zeae using a molecular phylogenetic analysis with multiple-gene genealogies. They discovered seven lineages within 37 isolates of G. zeae from a worldwide collection and demonstrated that the trichothecene chemotypes were not lineage specific. However, no Tri7 polymorphism was found in the Korean NIV-producing isolates in this study. These results are consistent with the hypothesis that there is a NIV chemotype-specific lineage in Korea, most likely lineage 6. In addition, the two Korean G. zeae chemotypes appear to be different lineages; 88-1 and H-11 were identified as lineages 6 and 7, respectively, based on sequence comparisons of Tri101. It is unclear whether the other Korean DON-producing isolates are lineage 7, as found for North American isolates. To confirm these results further studies are needed to determine the lineage(s) of Korean G. zeae isolates.
Differentiating chemotypes of G. zeae isolates is of great practical importance in several Asian countries, including Korea and Japan, where both chemotypes are common (15, 35, 36). Such a technique would also be useful in plant quarantine for North American countries, including the United States and Canada, where no NIV chemotypes have been detected (1, 27). The use of a PCR assay with Tri7 primers would avoid the difficulties that arise from time-consuming and laborious chemical analyses of toxins. In this study, we demonstrated that a PCR assay with Tri7 primers provides a rapid and reliable method for differentiating the two chemotypes of G. zeae from Korea and North America. If this PCR assay works on isolates from other locations, then it may be globally applicable. This assay also could be applicable for evaluating contamination of cereal samples with either DON- or NIV-producing isolates of G. zeae. This method, which is based on the size variation of PCR products, should be more reliable than other PCR-based detection systems that are based on the absence or presence of a PCR band and which may yield false-negative results. In addition, our studies of the nucleotide sequences of the Tri genes from the two chemotypes will allow development of a DNA probe hybridizing to different restriction fragment(s) of genomic DNA from each chemotype.
The sequence similarity in other Tri genes and noncoding regions between the two chemotypes of G. zeae most likely reflects their evolutionary divergence. However, the conservation of mutations, except for the number of repeats, within the Tri7 genes of DON isolates from both Korea and the United States leads us to question whether all DON isolates in different lineages have a common ancestry. Future studies are needed to determine if the Tri7 polymorphisms found in this study also are found in the other lineages.
| |
ACKNOWLEDGMENTS |
|---|
This study was supported by special research funds from the Korean Ministry of Agriculture and Forestry for 2000-2001 and by a postdoctoral fellowship to T.L. and graduate fellowships to D.-W.O., J.L., and H.-S.K. from the Korean Ministry of Education through the Brain Korea 21 Project.
We thank Robert L. Bowden of the Department of Plant Pathology, Kansas State University, Manhattan, for providing G. zeae strains for this study.
| |
FOOTNOTES |
|---|
* Corresponding author. Mailing address: School of Agricultural Biotechnology, Seoul National University, Suwon 441-744, Korea. Phone: 82-31-290-2443. Fax: 82-31-294-5881. E-mail: lee2443{at}snu.ac.kr.
| |
REFERENCES |
|---|
|
Top
Abstract
Introduction
Materials and Methods
Results
Discussion
References
|
|---|
| 1. | Abbas, H. K., C. J. Mirocha, T. Kommedahl, R. F. Vesonder, and P. Golinski. 1989. Production of trichothecene and non-trichothecene mycotoxins by Fusarium species isolated from maize in Minnesota. Mycopathologia 108:55-58[CrossRef][Medline]. |
| 2. |
Alexander, N. J.,
T. M. Hohn, and S. P. McCormick.
1988.
The TRI11 gene of Fusarium sporotrichioides encodes a cytochrome P-450 monooxygenase required for C-15 hydroxylation in trichothecene biosynthesis.
Appl. Environ. Microbiol.
64:221-225 |
| 3. | Alexander, N. J., S. P. McCormick, and T. M. Hohn. 1999. TRI12, a trichothecene efflux pump from Fusarium sporotrichioides: gene isolation and expression in yeast. Mol. Gen. Genet. 261:977-984[CrossRef][Medline]. |
| 4. | Altschul, S. F., W. Gish, W. Miller, E. W. Myers, and D. J. Lipman. 1990. Basic local alignment search tool. J. Mol. Biol. 215:403-410[CrossRef][Medline]. |
| 5. | Bowden, R. L., and J. F. Leslie. 1999. Sexual recombination in Gibberella zeae. Phytopathology 89:182-188[Medline]. |
| 6. | Brown, D. W., S. P. McCormick, N. J. Alexander, R. H. Proctor, and A. E. Desjardins. 2001. A genomic and biochemical approach to trichothecene diversity in Fusarium sporotrichioides and Fusarium graminearum. Fungal Genet. Newsl. 48(Suppl.):165. |
| 7. | Cook, R. J. 1981. Fusarium diseases of wheat and other small grains in North America, p. 39-52. In P. E. Nelson, and T. A. Toussoun (ed.), Fusarium diseases, biology, and taxonomy. The Pennsylvania State University Press, University Park. |
| 8. | Correll, J. C., C. J. R. Klittich, and J. F. Leslic. 1987. Nitrate nonutilizing mutants and their use in vegetative compatibility tests. Phytopathology 77:1640-1646[CrossRef]. |
| 9. |
Desjardins, A. E.,
T. M. Hohn, and S. P. McCormick.
1993.
Trichothecene biosynthesis in Fusarium species: chemistry, genetics, and significance.
Microbiol. Rev.
57:595-604 |
| 10. | Desjardins, A. E., H. K. Manadhar, R. D. Plattner, C. M. Maragos, K. Shrestha, and S. P. McCormick. 2000. Occurrence of Fusarium species and mycotoxins in Nepalese maize and wheat and the effect of traditional processing methods on mycotoxin levels. J. Agric. Food Chem. 48:1377-1383[CrossRef][Medline]. |
| 11. | Hohn, T. M., and P. D. Beremand. 1989. Isolation and nucleotide sequence of a sesquiterpene cyclase gene from trichothecene-producing fungus Fusarium sporotrichioides. Gene 79:131-138[CrossRef][Medline]. |
| 12. | Hohn, T. M., A. E. Desjardins, and S. P. McCormick. 1995. The Tri4 gene of Fusarium sporotrichioides encodes a cytochrome P450 monooxygenase involved in trichothecene biosynthesis. Mol. Gen. Genet. 248:95-102[CrossRef][Medline]. |
| 13. | Hohn, T. M., S. P. McCormick, N. J. Alexander, A. E. Desjardins, and R. H. Proctor. 1998. Function and biosynthesis of trichothecenes produced by Fusarium species, p. 17-24. In K. Kohmoto, and O. C. Yoder (ed.), Molecular genetics of host-specific toxins in plant diseases. Kluwer Academic Publishers, Dordrecht, The Netherlands. |
| 14. | Ichinoe, M., R. Amano, N. Morooka, T. Yoshizawa, T. Suzuki, and M. Kurisu. 1980. Geographic difference of toxigenic fungi of Fusarium species. Proc. Jpn. Assoc. Mycotoxicol. 11:20-22. |
| 15. |
Ichinoe, M.,
H. Kurata,
Y. Sugiura, and Y. Ueno.
1983.
Chemotaxonomy of Gibberella zeae with special reference to production of trichothecenes and zearalenone.
Appl. Environ. Microbiol.
46:1364-1369 |
| 16. |
Kerenyi, Z.,
K. Zeller,
L. Hornok, and J. F. Leslie.
1999.
Molecular standardization of mating type terminology in the Gibberella fujikuroi species complex.
Appl. Environ. Microbiol.
65:4071-4076 |
| 17. |
Kim, J.-C.,
H.-J. Kang,
D.-H. Lee,
Y.-W. Lee, and T. Yoshizawa.
1993.
Natural occurrence of Fusarium mycotoxins (trichothecenes and zearalenone) in barley and corn in Korea.
Appl. Environ. Microbiol.
59:3798-3802 |
| 18. |
Kimura, M.,
I. Kaneko,
M. Komiyama,
A. Takatsuki,
H. Koshino,
K. Yoneyama, and I. Yamaguchi.
1998.
Trichothecene 3-O-acetyltransferase protects both the producing organism and transformed yeast from related mycotoxins. Cloning and characterization of Tri101.
J. Biol. Chem.
273:1654-1661 |
| 19. | Kimura, M., G. Matsumoto, Y. Shingu, K. Yoneyama, and I. Yamaguchi. 1998. The mystery of the trichothecene 3-O-acetyltransferase gene. Analysis of the region around Tri101 and characterization of its homologue from Fusarium sporotrichioides. FEBS Lett. 435:163-168[CrossRef][Medline]. |
| 20. | Kommedahl, T., and C. E. Windels. 1981. Root-, stalk-, and ear-infecting Fusarium species on corn in the USA, p. 94-103. In P. E. Nelson, T. A. Toussoun, and R. J. Cook (ed.), Fusarium diseases, biology, and taxonomy. The Pennsylvania State University Press, University Park. |
| 21. | Liu, Z., and N. C. Mishra. 1995. A single-tube method for plasmid mini-prep from large numbers of clones for direct screening by size or restriction digestion. BioTechniques 18:214-217[Medline]. |
| 22. | Logrieco, A., A. Bottalico, and C. Altomare. 1988. Chemotaxonomic observation on zearalenone and trichothecene production by Gibberella zeae from cereals in southern Italy. Mycologia 80:892-895. |
| 23. | Marasas, W. F. O., P. E. Nelson, and T. A. Toussoun. 1984. Toxigenic Fusarium species: identity and mycotoxicology. The Pennsylvania State University Press, University Park. |
| 24. | Matsumoto, G., J. Wuchiyama, Y. Shingu, M. Kimura, K. Yoneyama, and I. Yamaguchi. 1999. The trichothecene biosynthesis regulating gene from the type B producer Fusarium strains: sequence of Tri6 and its expression in Escherichia coli. Biosci. Biotechnol. Biochem. 63:2001-2004[CrossRef][Medline]. |
| 25. | McCormick, S. P., T. M. Hohn, and A. E. Desjardins. 1996. Isolation and characterization of Tri3, a gene encoding 15-O-acetyltransferase from Fusarium sporotrichioides. Appl. Environ. Microbiol. 62:353-359[Abstract]. |
| 26. | McCormick, S. P., T. M. Hohn, A. E. Desjardins, R. H. Proctor, and N. J. Alexander. 1998. Role of toxins in plant microbial interactions, p. 17-30. In J. T. Romeo, K. R. Downum, and R. Verpoorte (ed.), Recent advances in phytochemistry, vol. 32. Phytochemical signals and plant-microbe interactions. Plenum Press, New York, n.y. |
| 27. |
Mirocha, C. J.,
H. K. Abbas,
C. E. Windels, and W. Xie.
1989.
Variation in deoxynivalenol, 15-acetyldeoxynivalenol, 3-acetyldeoxynivalenol, and zearalenone production by Fusarium graminearum isolates.
Appl. Environ. Microbiol.
55:1315-1316 |
| 28. | Nelson, P. E., B. W. Pennypacker, T. A. Toussoun, and R. K. Horst. 1975. Fusarium stub dicback of carnation. Phytopathology 65:575-581. |
| 29. |
Ochman, H.,
A. S. Gerber, and D. L. Hartl.
1988.
Genetic applications of an inverse polymerase chain reaction.
Genetics
120:621-624 |
| 30. |
O'Donnell, K.,
H. C. Kistler,
B. K. Tacke, and H. H. Casper.
2000.
Gene genealogies reveal global phylogeographic structure and reproductive isolation among lineages of Fusarium graminearum, the fungus causing wheat scab.
Proc. Natl. Acad. Sci. USA
97:7905-7910 |
| 31. | Proctor, R. H., T. M. Hohn, and S. P. McCormick. 1995. Reduced virulence of Gibberella zeae caused by disruption of a trichothecene toxin biosynthetic gene. Mol. Plant-Microbe Interact. 8:593-601[Medline]. |
| 32. | Proctor, R. H., T. M. Hohn, S. P. McCormick, and A. E. Desjardins. 1995. Tri6 encodes an unusual zinc finger protein involved in regulation of trichothecene biosynthesis in Fusarium sporotrichioides. Appl. Environ. Microbiol. 61:1923-1930[Abstract]. |
| 33. | Ryu, J. C., K. Ohtsubo, N. Izumiyama, K. Nakamura, T. Tanaka, H. Yamamura, and Y. Ueno. 1988. The acute and chronic toxicities of nivalenol in mice. Fund. Appl. Toxicol. 11:38-47[CrossRef][Medline]. |
| 34. | Sambrook, J., E. F. Fritsch, and T. Maniatis. 1989. Molecular cloning: a laboratory manual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. |
| 35. | Seo, J.-A., J.-C. Kim, D.-H. Lee, and Y.-W. Lee. 1996. Variation in 8-ketotrichothecenes and zearalenone production by Fusarium graminearum isolates from corn and barley in Korea. Mycopathologia 134:31-37[CrossRef]. |
| 36. |
Sugiura, Y.,
Y. Watanabe,
T. Tanaka,
S. Yamamoto, and Y. Ueno.
1990.
Occurrence of Gibberella zeae strains that produce both nivalenol and deoxynivalenol.
Appl. Environ. Microbiol.
56:3047-3051 |
| 37. | Sydenham, E. W., W. F. O. Marasas, P. G. Thiel, G. S. Shephard, and J. J. Nieuwenhuis. 1991. Production of mycotoxins by selected Fusarium graminearum and F. crookwellense isolates. Food Addit. Contam. 8:31-41[Medline]. |
| 38. | Tanaka, T., A. Hasegawa, S. Yamamoto, U. S. Lee, Y. Sugiura, and Y. Ueno. 1988. Worldwide contamination of cereals by the Fusarium mycotoxins nivalenol, deoxynivalenol and zearalenone. I. Survey of 19 countries. J. Agric. Food Chem. 36:979-983[CrossRef]. |
| 39. | Wuchiyama, J., M. Kimura, and I. Yamaguchi. 2000. A trichothecene efflux pump encoded by Tri102 in the biosynthesis gene cluster of Fusarium graminearum. J. Antibiot. 53:196-200[Medline]. |
| 40. | Yoshizawa, T., and Y. Z. Jin. 1995. Natural occurrence of acetylated derivatives of deoxynivalenol and nivalenol in wheat and barley in Japan. Food Addit. Contam. 12:689-694[Medline]. |
| 41. | Yun, S.-H., T. Arie, I. Kaneko, O. C. Yoder, and B. G. Turgeon. 2000. Molecular organization of mating type loci in heterothallic, homothallic, and asexual Gibberella/Fusarium species. Fungal Genet. Biol. 31:7-20[CrossRef][Medline]. |
Applied and Environmental Microbiology, July 2001, p. 2966-2972, Vol. 67, No. 7
0099-2240/01/$04.00+0 DOI: 10.1128/AEM.67.7.2966-2972.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
This article has been cited by other articles:
-
Lee, J., Chang, I.-Y., Kim, H., Yun, S.-H., Leslie, J. F., Lee, Y.-W.
(2009). Genetic Diversity and Fitness of Fusarium graminearum Populations from Rice in Korea. Appl. Environ. Microbiol.
75: 3289-3295
[Abstract] [Full Text] -
Tokai, T., Fujimura, M., Inoue, H., Aoki, T., Ohta, K., Shibata, T., Yamaguchi, I., Kimura, M.
(2005). Concordant evolution of trichothecene 3-O-acetyltransferase and an rDNA species phylogeny of trichothecene-producing and non-producing fusaria and other ascomycetous fungi. Microbiology
151: 509-519
[Abstract] [Full Text] -
McCormick, S. P., Harris, L. J., Alexander, N. J., Ouellet, T., Saparno, A., Allard, S., Desjardins, A. E.
(2004). Tri1 in Fusarium graminearum Encodes a P450 Oxygenase. Appl. Environ. Microbiol.
70: 2044-2051
[Abstract] [Full Text] -
Kimura, M., Tokai, T., Matsumoto, G., Fujimura, M., Hamamoto, H., Yoneyama, K., Shibata, T., Yamaguchi, I.
(2003). Trichothecene Nonproducer Gibberella Species Have Both Functional and Nonfunctional 3-O-Acetyltransferase Genes. Genetics
163: 677-684
[Abstract] [Full Text] -
Bakan, B., Giraud-Delville, C., Pinson, L., Richard-Molard, D., Fournier, E., Brygoo, Y.
(2002). Identification by PCR of Fusarium culmorum Strains Producing Large and Small Amounts of Deoxynivalenol. Appl. Environ. Microbiol.
68: 5472-5479
[Abstract] [Full Text] -
McCormick, S. P., Alexander, N. J.
(2002). Fusarium Tri8 Encodes a Trichothecene C-3 Esterase. Appl. Environ. Microbiol.
68: 2959-2964
[Abstract] [Full Text] -
Lee, T., Han, Y.-K., Kim, K.-H., Yun, S.-H., Lee, Y.-W.
(2002). Tri13 and Tri7 Determine Deoxynivalenol- and Nivalenol-Producing Chemotypes of Gibberella zeae. Appl. Environ. Microbiol.
68: 2148-2154
[Abstract] [Full Text] -
Jurgenson, J. E., Bowden, R. L., Zeller, K. A., Leslie, J. F., Alexander, N. J., Plattner, R. D.
(2002). A Genetic Map of Gibberella zeae (Fusarium graminearum). Genetics
160: 1451-1460
[Abstract] [Full Text]
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Copyright © 2009 by the American Society for Microbiology. For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»