The evolution of fungicide resistance.

Fungicides are widely used in developed agricultural systems to control disease and safeguard crop yield and quality. Over time, however, resistance to many of the most effective fungicides has emerged and spread in pathogen populations, compromising disease control. This review describes the development of resistance using case histories based on four important diseases of temperate cereal crops: eyespot (Oculimacula yallundae and Oculimacula acuformis), Septoria tritici blotch (Zymoseptoria tritici), powdery mildew (Blumeria graminis), and Fusarium ear blight (a complex of Fusarium and Microdochium spp). The sequential emergence of variant genotypes of these pathogens with reduced sensitivity to the most active single-site fungicides, methyl benzimidazole carbamates, demethylation inhibitors, quinone outside inhibitors, and succinate dehydrogenase inhibitors illustrates an ongoing evolutionary process in response to the introduction and use of different chemical classes. Analysis of the molecular mechanisms and genetic basis of resistance has provided more rapid and precise methods for detecting and monitoring the incidence of resistance in field populations, but when or where resistance will occur remains difficult to predict. The extent to which the predictability of resistance evolution can be improved by laboratory mutagenesis studies and fitness measurements, comparison between pathogens, and reconstruction of evolutionary pathways is discussed. Risk models based on fungal life cycles, fungicide properties, and exposure to the fungicide are now being refined to take account of additional traits associated with the rate of pathogen evolution. Experimental data on the selection of specific mutations or resistant genotypes in pathogen populations in response to fungicide treatments can be used in models evaluating the most effective strategies for reducing or preventing resistance. Resistance management based on robust scientific evidence is vital to prolong the effective life of fungicides and safeguard their future use in crop protection.

[1]  N. Hawkins,et al.  Constraints on the evolution of azole resistance in plant pathogenic fungi , 2013 .

[2]  H. Sierotzki,et al.  A review of current knowledge of resistance aspects for the next-generation succinate dehydrogenase inhibitor fungicides. , 2013, Phytopathology.

[3]  S. Powers,et al.  Evaluation of a matrix to calculate fungicide resistance risk. , 2014, Pest management science.

[4]  D. Kelly,et al.  Impact of Recently Emerged Sterol 14α-Demethylase (CYP51) Variants of Mycosphaerella graminicola on Azole Fungicide Sensitivity , 2011, Applied and Environmental Microbiology.

[5]  H. Sierotzki,et al.  Point Mutation in Cytochrome b Gene Conferring Resistance to Strobilurin Fungicides in Erysiphe graminis f. sp. tritici Field Isolates , 2000 .

[6]  J. West,et al.  Impacts of changing climate and agronomic factors on fusarium ear blight of wheat in the UK , 2012 .

[7]  B. McDonald,et al.  Frequency of mutations associated with fungicide resistance and population structure of Mycosphaerella graminicola in Tunisia , 2011, European Journal of Plant Pathology.

[8]  S. Welham,et al.  Changes in populations of the eyespot fungi Tapesia yallundae and T. acuformis under different fungicide regimes in successive crops of winter wheat, 1984–2000 , 2002 .

[9]  J. Horsfall Fungicides and their action , 1946 .

[10]  E. M. Cooper,et al.  Systematic mutational analysis of the yeast beta-tubulin gene. , 1994, Molecular biology of the cell.

[11]  K. Hammond-Kosack,et al.  Exploitation of genomics in fungicide research: current status and future perspectives. , 2013, Molecular plant pathology.

[12]  L. Hoffmann,et al.  Differences between the succinate dehydrogenase sequences of isopyrazam sensitive Zymoseptoria tritici and insensitive Fusarium graminearum strains. , 2013, Pesticide Biochemistry and Physiology.

[13]  M. Shaw,et al.  Sterol content analysis suggests altered eburicol 14alpha-demethylase (CYP51) activity in isolates of Mycosphaerella graminicola adapted to azole fungicides. , 2009, FEMS microbiology letters.

[14]  M. Milgroom A Simulation Analysis of the Epidemiological Principles for Fungicide Resistance Management in Pathogen Populations , 1988 .

[15]  T. Miles,et al.  Screening and characterization of resistance to succinate dehydrogenase inhibitors in Alternaria solani , 2014 .

[16]  R. Becher,et al.  Fungal cytochrome P450 sterol 14α-demethylase (CYP51) and azole resistance in plant and human pathogens , 2012, Applied Microbiology and Biotechnology.

[17]  A. Gautier,et al.  Mutations in the CYP51 gene correlated with changes in sensitivity to sterol 14 alpha-demethylation inhibitors in field isolates of Mycosphaerella graminicola. , 2007, Pest management science.

[18]  P. Karlovsky,et al.  Adaptation of Fusarium graminearum to tebuconazole yielded descendants diverging for levels of fitness, fungicide resistance, virulence, and mycotoxin production. , 2010, Phytopathology.

[19]  E. Mellado,et al.  Targeted Gene Disruption of the 14-α Sterol Demethylase (cyp51A) in Aspergillus fumigatus and Its Role in Azole Drug Susceptibility , 2005, Antimicrobial Agents and Chemotherapy.

[20]  F. Delmotte,et al.  At Least Two Origins of Fungicide Resistance in Grapevine Downy Mildew Populations , 2007, Applied and Environmental Microbiology.

[21]  S. Fillinger,et al.  Fitness measurement reveals contrasting costs in homologous recombinant mutants of Botrytis cinerea resistant to succinate dehydrogenase inhibitors. , 2014, Fungal genetics and biology : FG & B.

[22]  J. Vontas,et al.  Biological and molecular characterization of laboratory mutants of Cercospora beticola resistant to Qo inhibitors , 2006, European Journal of Plant Pathology.

[23]  S. Foster,et al.  Amplification of a Cytochrome P450 Gene Is Associated with Resistance to Neonicotinoid Insecticides in the Aphid Myzus persicae , 2010, PLoS genetics.

[24]  S. Dutzmann,et al.  JAU 6476 - a new dimension DMI fungicide. , 2002 .

[25]  F. C. van den Bosch,et al.  Delaying selection for fungicide insensitivity by mixing fungicides at a low and high risk of resistance development: a modeling analysis. , 2011, Phytopathology.

[26]  C. Mundt,et al.  First Report of Resistance to QoI Fungicides in North American Populations of Zymoseptoria tritici, Causal Agent of Septoria Tritici Blotch of Wheat. , 2013, Plant disease.

[27]  M. Pfrender,et al.  Rapid evolution in response to introduced predators I: rates and patterns of morphological and life-history trait divergence , 2007, BMC Evolutionary Biology.

[28]  M. Shaw,et al.  Sensitivity distributions and cross-resistance patterns of Mycosphaerella graminicola to fluquinconazole, prochloraz and azoxystrobin over a period of 9 years , 2005 .

[29]  J. Lucas,et al.  Foresight project on global food and farming futures: advances in plant disease and pest management , 2011 .

[30]  H. Fehrmann,et al.  Fungicide resistance of Septoria nodorum and Pseudocercosporella herpotrichoides I. Effect of fungicide application on the frequency of resistant spores in the field. , 1980 .

[31]  Rory Hillocks,et al.  Farming with fewer pesticides: EU pesticide review and resulting challenges for UK agriculture , 2012 .

[32]  P. Dyer,et al.  Cloning of the CYP51 gene from the eyespot pathogen Tapesia yallundae indicates that resistance to the DMI fungicide prochloraz is not related to sequence changes in the gene encoding the target site enzyme. , 2001, FEMS microbiology letters.

[33]  G. Schwarz,et al.  CAPS and DHPLC Analysis of a Single Nucleotide Polymorphism in the Cytochrome b Gene Conferring Resistance to Strobilurins in Field Isolates of Blumeria graminis f. sp. hordei , 2003 .

[34]  A. C. King First record of Tapesia yallundae as the teleomorph of Pseudocercosporella herpotrichoides var. acuformis, and its occurrence in the field in the Federal Republic of Germany , 1990 .

[35]  M. Wolfe Trying to understand and control powdery mildew , 1984 .

[36]  P. E. Russell,et al.  Sensitivity of CYP51 genotypes to DMI fungicides in mycosphaerella graminicola. , 2008 .

[37]  S. Powers,et al.  Fungicide resistance risk assessment based on traits associated with the rate of pathogen evolution. , 2015, Pest management science.

[38]  P. E. Russell A century of fungicide evolution , 2005, The Journal of Agricultural Science.

[39]  K. Downing,et al.  Meiosis-specific failure of cell cycle progression in fission yeast by mutation of a conserved beta-tubulin residue. , 2003, Molecular biology of the cell.

[40]  C. Gilligan,et al.  Invasion thresholds for fungicide resistance: deterministic and stochastic analyses , 1999, Proceedings of the Royal Society of London. Series B: Biological Sciences.

[41]  J. Oakeshott,et al.  Evolution in agriculture: the application of evolutionary approaches to the management of biotic interactions in agro-ecosystems , 2011, Evolutionary applications.

[42]  Hee-won Park,et al.  Structural basis of human CYP51 inhibition by antifungal azoles. , 2010, Journal of molecular biology.

[43]  F. Trail,et al.  For Blighted Waves of Grain: Fusarium graminearum in the Postgenomics Era , 2009, Plant Physiology.

[44]  E. Mullins,et al.  Pyraclostrobin reduces germ tube growth of QoI-resistant Mycosphaerella graminicola pycnidiospores and the severity of septoria tritici blotch on winter wheat , 2010 .

[45]  P. Leroux,et al.  Mutations of the β-tubulin gene associated with different phenotypes of benzimidazole resistance in the cereal eyespot fungi Tapesia yallundae and Tapesia acuformis. , 1999 .

[46]  恵子 鈴木,et al.  ベンズイミダゾール系殺菌剤および methyl N-(3, 5-dichlorophenyl) carbamate 間における負相関交差耐性 , 1984 .

[47]  S. Freeman,et al.  Identification and Characterization of Benomyl-Resistant and -Sensitive Populations of Colletotrichum gloeosporioides from Statice (Limonium spp.). , 2006, Phytopathology.

[48]  H. Fehrmann,et al.  Five years' results from a long-term field experiment on carbendazim resistance of Pseudocercosporella herpotrichoides (Fron) Deighton , 1982 .

[49]  S. Powles,et al.  Evolution in action: plants resistant to herbicides. , 2010, Annual review of plant biology.

[50]  J. Bartlett,et al.  The epidemic of antibiotic-resistant infections: a call to action for the medical community from the Infectious Diseases Society of America. , 2008, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[51]  J. Brown,et al.  Sequence variation in the CYP51 gene of Blumeria graminis associated with resistance to sterol demethylase inhibiting fungicides. , 2005, Fungal genetics and biology : FG & B.

[52]  P. Karlovsky,et al.  Identification of ABC Transporter Genes of Fusarium graminearum with Roles in Azole Tolerance and/or Virulence , 2013, PloS one.

[53]  Pari Skamnioti,et al.  Host perception and signal transduction studies in wild-type Blumeria graminis f. sp. hordei and a quinoxyfen-resistant mutant implicate quinoxyfen in the inhibition of serine esterase activity. , 2008, Pest management science.

[54]  D. Denning,et al.  Environmental fungicides and triazole resistance in Aspergillus. , 2014, Pest management science.

[55]  N. Paveley,et al.  Derivation and testing of a model to predict selection for fungicide resistance , 2011 .

[56]  D. Kelly,et al.  Heterologous Expression of Mutated Eburicol 14α-Demethylase (CYP51) Proteins of Mycosphaerella graminicola To Assess Effects on Azole Fungicide Sensitivity and Intrinsic Protein Function , 2010, Applied and Environmental Microbiology.

[57]  Zhonghua Ma,et al.  Characterization and PCR-based detection of benzimidazole-resistant isolates of Monilinia laxa in California. , 2005, Pest management science.

[58]  S. Katiyar,et al.  Site‐directed mutagenesis of Saccharomyces cerevisiae β‐tubulin: interaction between residue 167 and benzimidazole compounds , 1996, FEBS letters.

[59]  Pedro W. Crous,et al.  Eyespot of Cereals Revisited: ITS phylogeny Reveals New Species Relationships , 2003, European Journal of Plant Pathology.

[60]  P. Leroux,et al.  Caractérisation des souches de Pseudocercosporella herpotrichoides, agent du piétin-verse des céréales, résistantes au prochloraze, isolées en France sur blé tendre d'hiver , 1991 .

[61]  S. Kelly,et al.  Characterization of the sterol 14α-demethylases of Fusarium graminearum identifies a novel genus-specific CYP51 function. , 2013, The New phytologist.

[62]  M Lappé,et al.  Genetic control. , 1972, The New England journal of medicine.

[63]  R. P. White,et al.  Climate change increases risk of fusarium ear blight on wheat in central China , 2014 .

[64]  H. Hamamoto,et al.  Tandem Repeat of a Transcriptional Enhancer Upstream of the Sterol 14α-Demethylase Gene (CYP51) inPenicillium digitatum , 2000, Applied and Environmental Microbiology.

[65]  S. Short,et al.  Amino acid substitutions in the CytR repressor which alter its capacity to regulate gene expression , 1992, Journal of bacteriology.

[66]  M. Bolton,et al.  Characterization of cytochrome b from European field isolates of Cercospora beticola with quinone outside inhibitor resistance , 2012, European Journal of Plant Pathology.

[67]  K. Kuchler,et al.  Mechanisms of resistance to azole antifungal agents in Candida albicans isolates from AIDS patients involve specific multidrug transporters , 1995, Antimicrobial agents and chemotherapy.

[68]  Dominique Sanglard,et al.  Amino Acid Substitutions in the Cytochrome P-450 Lanosterol 14α-Demethylase (CYP51A1) from Azole-Resistant Candida albicans Clinical Isolates Contribute to Resistance to Azole Antifungal Agents , 1998, Antimicrobial Agents and Chemotherapy.

[69]  M. Fraaije,et al.  Risk assessment studies on succinate dehydrogenase inhibitors, the new weapons in the battle to control Septoria leaf blotch in wheat. , 2012, Molecular plant pathology.

[70]  M. J. Griffin,et al.  Survey of benomyl resistance in Pseudocercosporella herpotrichoides on winter wheat and barley in England and Wales in 1983 , 1985 .

[71]  R. Dean,et al.  The evolutionary history of Cytochrome P450 genes in four filamentous Ascomycetes , 2007, BMC Evolutionary Biology.

[72]  Jian-xin Wang,et al.  Characterization and fitness of carbendazim-resistant strains of Fusarium graminearum (wheat scab). , 2007, Pest Management Science.

[73]  O. Yarden Mutations leading to substitutions at amino acids 198 and 200 of beta-tubulin that correlate with benomyl-resistance phenotypes of field strains of Botrytis cinerea , 1993 .

[74]  P. Leroux,et al.  Fungicide resistance status in French populations of the wheat eyespot fungi Oculimacula acuformis and Oculimacula yallundae. , 2013, Pest management science.

[75]  M. Shaw,et al.  Wheat archive links long-term fungal pathogen population dynamics to air pollution. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[76]  M. Noble,et al.  RESISTANCE TO MERCURY OF PYRENOPHORA AVENAE IN SCOTTISH SEED OATS , 1966 .

[77]  W. Melchers,et al.  A New Aspergillus fumigatus Resistance Mechanism Conferring In Vitro Cross-Resistance to Azole Antifungals Involves a Combination of cyp51A Alterations , 2007, Antimicrobial Agents and Chemotherapy.

[78]  B. Oakley,et al.  Identification of an amino acid substitution in the benA, beta-tubulin gene of Aspergillus nidulans that confers thiabendazole resistance and benomyl supersensitivity. , 1990, Cell motility and the cytoskeleton.

[79]  G. B. Golding,et al.  Antibiotic resistance is ancient , 2011, Nature.

[80]  H. Inoue,et al.  Sensitivity of Neurospora crassa to benzimidazoles and N-phenylcarbamates : effect of amino acid substitutions at position 198 in β-tubulin , 1992 .

[81]  M. Milgroom,et al.  Inheritance of triadimenol resistance in Pyrenophora teres , 1992 .

[82]  B. McDonald,et al.  Evolution of the CYP51 gene in Mycosphaerella graminicola: evidence for intragenic recombination and selective replacement. , 2008, Molecular plant pathology.

[83]  Zhonghua Ma,et al.  Advances in understanding molecular mechanisms of fungicide resistance and molecular detection of resistant genotypes in phytopathogenic fungi , 2005 .

[84]  F. Burnett,et al.  Sensitivity of powdery mildew and yellow rust to DMI, morpholine and strobilurin fungicides in England and Scotland. , 2000 .

[85]  D. Livermore Has the era of untreatable infections arrived? , 2009, The Journal of antimicrobial chemotherapy.

[86]  S. Bonhoeffer,et al.  Can high-risk fungicides be used in mixtures without selecting for fungicide resistance? , 2013, Phytopathology.

[87]  L. Panella,et al.  Analysis of β-tubulin Gene Fragments from Benzimidazole-sensitive and -tolerant Cercospora beticola , 2006 .

[88]  B. Oakley,et al.  Amino acid alterations in the benA (beta-tubulin) gene of Aspergillus nidulans that confer benomyl resistance. , 1992, Cell motility and the cytoskeleton.

[89]  Zhonghua Ma,et al.  Identification and Characterization of Benzimidazole Resistance in Monilinia fructicola from Stone Fruit Orchards in California , 2003, Applied and Environmental Microbiology.

[90]  Zhonghua Ma,et al.  Detection and dynamics of different carbendazim-resistance conferring β-tubulin variants of Gibberella zeae collected from infected wheat heads and rice stubble in China. , 2014, Pest management science.

[91]  M. Waterman,et al.  Crystal Structures of Trypanosoma brucei Sterol 14α-Demethylase and Implications for Selective Treatment of Human Infections*♦ , 2009, The Journal of Biological Chemistry.

[92]  J. Brown,et al.  Isogamous, hermaphroditic inheritance of mitochondrion-encoded resistance to Qo inhibitor fungicides in Blumeria graminis f. sp. tritici. , 2002, Fungal genetics and biology : FG & B.

[93]  H. Sierotzki,et al.  Cytochrome b gene sequence and structure of Pyrenophora teres and P. tritici-repentis and implications for QoI resistance. , 2007, Pest management science.

[94]  Y. Yin,et al.  Characterization of sterol demethylation inhibitor-resistant isolates of Fusarium asiaticum and F. graminearum collected from wheat in China. , 2009, Phytopathology.

[95]  Roger Jones,et al.  Scab of Wheat and Barley: A Re-emerging Disease of Devastating Impact. , 1997, Plant disease.

[96]  H. Inoue,et al.  Amino-acid alterations in the β-tubilin gene of Neurospora crassa that confer resistance to carbendazim and diethofencarb , 1994, Current Genetics.

[97]  H. Sierotzki,et al.  Mode of resistance to respiration inhibitors at the cytochrome bc1 enzyme complex of Mycosphaerella fijiensis field isolates , 2000 .

[98]  M. Farman,et al.  Field Resistance to Strobilurin (Q(o)I) Fungicides in Pyricularia grisea Caused by Mutations in the Mitochondrial Cytochrome b Gene. , 2003, Phytopathology.

[99]  M. Coffey,et al.  Biodegradation of metalaxyl in avocado soils , 1985 .

[100]  K. Gallimore,et al.  Sensitivity of Pseudoeevcosporella herpotriehoides to the fungicide prochloraz , 1987 .

[101]  L. Hoffmann,et al.  Evidence for natural resistance towards trifloxystrobin in Fusarium graminearum , 2011, European Journal of Plant Pathology.

[102]  R. Cook,et al.  Disease-induced losses in winter wheat in England and Wales 1985–1989 , 1991 .

[103]  J. Vontas,et al.  Effect of DMI-resistance mechanisms on cross-resistance patterns, fitness parameters and aflatoxin production in Aspergillus parasiticus Speare. , 2012, Fungal genetics and biology : FG & B.

[104]  P. Burns,et al.  DNA sequence analysis of mutagenicity and site specificity of ethyl methanesulfonate in Uvr+ and UvrB- strains of Escherichia coli. , 1986, Genetics.

[105]  R. Rajendran,et al.  Azole Resistance of Aspergillus fumigatus Biofilms Is Partly Associated with Efflux Pump Activity , 2011, Antimicrobial Agents and Chemotherapy.

[106]  P. Dyer,et al.  Pathogenicity, host-specificity, and population biology of tapesia spp., causal agents of eyespot disease of cereals , 2000 .

[107]  C. Tanaka,et al.  Molecular analysis and characterization of the Cochliobolus heterostrophus beta-tubulin gene and its possible role in conferring resistance to benomyl. , 1998, The Journal of general and applied microbiology.

[108]  S. Georgopoulos,et al.  Genetic variability in the fungi and the problem of fungicide resistance , 1986 .

[109]  H. Sierotzki,et al.  Cytochrome b gene structure and consequences for resistance to Qo inhibitor fungicides in plant pathogens. , 2006, Pest management science.

[110]  U. Gisi Assessment of selection and resistance risk for demethylation inhibitor fungicides in Aspergillus fumigatus in agriculture and medicine: a critical review. , 2014, Pest management science.

[111]  W. Melchers,et al.  Azole Resistance Profile of Amino Acid Changes in Aspergillus fumigatus CYP51A Based on Protein Homology Modeling , 2010, Antimicrobial Agents and Chemotherapy.

[112]  M. McGrath Fungicide Resistance in Cucurbit Powdery Mildew: Experiences and Challenges. , 2001, Plant disease.

[113]  R. Nakaune,et al.  Benomyl resistance of Colletotrichum acutatum is caused by enhanced expression of beta-tubulin 1 gene regulated by putative leucine zipper protein CaBEN1. , 2007, Fungal genetics and biology : FG & B.

[114]  W. Uddin,et al.  Fitness and Competitive Ability of an Azoxystrobin-Resistant G143A Mutant of Magnaporthe oryzae from Perennial Ryegrass. , 2009, Plant disease.

[115]  James K. M. Brown,et al.  Selection on responses of barley powdery mildew to morpholine and piperidine fungicides , 1992 .

[116]  D. J. Royle,et al.  Airborne inoculum as a major source of Septoria tritici (Mycosphaerella graminicola) infections in winter wheat crops in the UK , 1989 .

[117]  N. Gudmestad,et al.  Effect of the F129L Mutation in Alternaria solani on Fungicides Affecting Mitochondrial Respiration. , 2005, Plant disease.

[118]  Shunji Suzuki,et al.  Development of a multiplex allele-specific primer PCR assay for simultaneous detection of QoI and CAA fungicide resistance alleles in Plasmopara viticola populations. , 2013, Pest management science.

[119]  Characterisation of benzimidazole resistance of Cercospora beticola in Serbia using PCR-based detection of resistance-associated mutations of the β-tubulin gene , 2013, European Journal of Plant Pathology.

[120]  J. Brownstein,et al.  Emerging fungal threats to animal, plant and ecosystem health , 2012, Nature.

[121]  Nicholas Fisher,et al.  Re-examination of inhibitor resistance conferred by Qo-site mutations in cytochrome b using yeast as a model system. , 2005, Pest management science.

[122]  C. Gilligan,et al.  Changes in fungicide sensitivity and relative species abundance in Oculimacula yallundae and O. acuformis populations (eyespot disease of cereals) in Western Europe , 2008 .

[123]  A. de Vicente,et al.  Mechanisms of resistance to QoI fungicides in phytopathogenic fungi. , 2008, International microbiology : the official journal of the Spanish Society for Microbiology.

[124]  A. Siah,et al.  Azoxystrobin resistance of French Mycosphaerella graminicola strains assessed by four in vitro bioassays and by screening of G143A substitution , 2010 .

[125]  R. Edwards,et al.  Key role for a glutathione transferase in multiple-herbicide resistance in grass weeds , 2013, Proceedings of the National Academy of Sciences.

[126]  E. M. Ponte,et al.  Sensitivity of Fusarium graminearum causing head blight of wheat in Brazil to tebuconazole and metconazole fungicides , 2012 .

[127]  L. Zwiers,et al.  ABC Transporters and Azole Susceptibility in Laboratory Strains of the Wheat Pathogen Mycosphaerella graminicola , 2002, Antimicrobial Agents and Chemotherapy.

[128]  L. Fraissinet-Tachet,et al.  Monitoring of Venturia inaequalis harbouring the QoI resistance G143A mutation in French orchards as revealed by PCR assays. , 2009, Pest management science.

[129]  P. E. Russell,et al.  Integrating Disease Control in Winter Wheat – Optimizing Fungicide Input , 2008 .

[130]  M. Huynen,et al.  Discovery of a hapE Mutation That Causes Azole Resistance in Aspergillus fumigatus through Whole Genome Sequencing and Sexual Crossing , 2012, PloS one.

[131]  T. S. Thind Fungicide resistance in crop protection: risk and management. , 2012 .

[132]  W. Köller,et al.  Fungal resistance to sterol biosynthesis inihitors: a new challenge , 1987 .

[133]  D. Hollomon,et al.  Combating plant diseases--the Darwin connection. , 2009, Pest management science.

[134]  D. Sanglard,et al.  Overexpression of the MDR1 Gene Is Sufficient To Confer Increased Resistance to Toxic Compounds in Candida albicans , 2006, Antimicrobial Agents and Chemotherapy.

[135]  R. Oliver,et al.  Governing principles can guide fungicide-resistance management tactics. , 2014, Annual review of phytopathology.

[136]  M. Brown,et al.  Carbendazim resistance in the eyespot pathogen Pseudocercosporella herpotrichoides , 1984 .

[137]  Paramvir S. Dehal,et al.  Finished Genome of the Fungal Wheat Pathogen Mycosphaerella graminicola Reveals Dispensome Structure, Chromosome Plasticity, and Stealth Pathogenesis , 2011, PLoS genetics.

[138]  W. Melchers,et al.  Academic Editor: Chris Kibbler, Royal Free Hospital London, United Kingdom , 2007 .

[139]  T. Michailides,et al.  Resistance to Boscalid Fungicide in Alternaria alternata Isolates from Pistachio in California. , 2007, Plant disease.

[140]  W. Köller,et al.  Characterization of spontaneous mutants of Magnaporthe grisea expressing stable resistance to the Qo-inhibiting fungicide azoxystrobin , 2003, Current Genetics.

[141]  R. Arango,et al.  Analysis of the CYP51 gene and encoded protein in propiconazole-resistant isolates of Mycosphaerella fijiensis. , 2009, Pest management science.

[142]  R. Oliver A reassessment of the risk of rust fungi developing resistance to fungicides. , 2014, Pest management science.

[143]  M. Dickman,et al.  Isolation of a beta-tubulin gene from Fusarium moniliforme that confers cold-sensitive benomyl resistance , 1996, Applied and environmental microbiology.

[144]  Pari Skamnioti,et al.  Genome Expansion and Gene Loss in Powdery Mildew Fungi Reveal Tradeoffs in Extreme Parasitism , 2010, Science.

[145]  T. Staub,et al.  Characterizing resistance risk of Erysiphe graminis f.sp. tritici to strobilurins , 2001 .

[146]  F. Burnett,et al.  QoI resistance development in populations of cereal pathogens in the UK. , 2003 .

[147]  S. Gurr,et al.  A single amino-acid substitution in the iron-sulphur protein subunit of succinate dehydrogenase determines resistance to carboxin in Mycosphaerella graminicola , 1998, Current Genetics.

[148]  P. E. Russell,et al.  The impact of the new European regulations on the management of crop diseases. , 2011 .

[149]  L. Zwiers,et al.  Control of Mycosphaerella graminicola on Wheat Seedlings by Medical Drugs Known To Modulate the Activity of ATP-Binding Cassette Transporters , 2007, Applied and Environmental Microbiology.

[150]  P. E. Russell,et al.  FRAC: combined resistance risk assessment , 2006 .

[151]  P. Dyer,et al.  Species and Mating-Type Distribution of Tapesia yallundae and T. acuformis and Occurrence of Apothecia in the U.S. Pacific Northwest. , 2002, Phytopathology.

[152]  L. Cowen,et al.  Evolution of Drug Resistance in Experimental Populations of Candida albicans , 2000, Journal of bacteriology.

[153]  G. Kema,et al.  MgMfs1, a major facilitator superfamily transporter from the fungal wheat pathogen Mycosphaerella graminicola, is a strong protectant against natural toxic compounds and fungicides. , 2007, Fungal genetics and biology : FG & B.

[154]  M. Dickman,et al.  Isolation, characterization, and expression of a second beta-tubulin-encoding gene from Colletotrichum gloeosporioides f. sp. aeschynomene , 1994, Applied and environmental microbiology.

[155]  P. Leroux,et al.  Polymorphism of 14α-demethylase Gene (CYP51) in the Cereal Eyespot Fungi Tapesia acuformis and Tapesia yallundae , 2003, European Journal of Plant Pathology.

[156]  T. Michailides,et al.  Progress in understanding molecular mechanisms and evolution of resistance to succinate dehydrogenase inhibiting (SDHI) fungicides in phytopathogenic fungi , 2010 .

[157]  Michael W. Shaw,et al.  The dose rate debate: Does the risk of fungicide resistance increase or decrease with dose? , 2011 .

[158]  D. Hollomon,et al.  Following the dynamics of strobilurin resistance in Blumeria graminis f.sp. tritici using quantitative allele-specific real-time PCR measurements with the fluorescent dye SYBR Green I , 2002 .

[159]  De Waard Resistance to fungicides which inhibit sterol 14_-demethylation, an historical perspective. , 1994 .

[160]  S. Somerville,et al.  Characterization of mutations in the beta-tubulin gene of benomyl-resistant field strains of Venturia inaequalis and other plant pathogenic fungi. , 1992 .

[161]  J. West,et al.  Role of Ascospores in Further Spread of QoI-Resistant Cytochrome b Alleles (G143A) in Field Populations of Mycosphaerella graminicola. , 2005, Phytopathology.

[162]  Christina A. Cuomo,et al.  The Fusarium graminearum Genome Reveals a Link Between Localized Polymorphism and Pathogen Specialization , 2007, Science.

[163]  N. Lima,et al.  Degradation of Metalaxyl and Folpet by Filamentous Fungi Isolated From Portuguese (Alentejo) Vineyard Soils , 2013, Archives of Environmental Contamination and Toxicology.

[164]  P. Dyer,et al.  Genetic Control of Resistance to the Sterol 14α-Demethylase Inhibitor Fungicide Prochloraz in the Cereal Eyespot PathogenTapesia yallundae , 2000, Applied and Environmental Microbiology.

[165]  D. Kelly,et al.  Molecular Modelling of the Emergence of Azole Resistance in Mycosphaerella graminicola , 2011, PloS one.

[166]  B. McDonald,et al.  QoI resistance emerged independently at least 4 times in European populations of Mycosphaerella graminicola. , 2009, Pest management science.

[167]  M. Kimura,et al.  Genotyping of benzimidazole-resistant and dicarboximide-resistant mutations in Botrytis cinerea using real-time polymerase chain reaction assays. , 2008, Phytopathology.

[168]  P. Dyer,et al.  Tapesia acuformis as a causal agent of eyespot disease of cereals and evidence for a heterothallic mating system using molecular markers , 1996 .

[169]  A. Julian,et al.  The induction and characterisation of isolates of Pseudocercosporella herpotrichoides with altered sensitivity to the fungicide prochloraz , 1994 .

[170]  M. Ogawa Antimicrobial Agents and Chemotherapy , 1964 .

[171]  M. Kimura,et al.  Characterization of QoI resistance in Botrytis cinerea and identification of two types of mitochondrial cytochrome b gene , 2009 .

[172]  F. C. van den Bosch,et al.  The Emergence of Resistance to Fungicides , 2014, PloS one.

[173]  M. Carstensen,et al.  Frequency of different CYP51-haplotypes of Mycosphaerella graminicola and their impact on epoxiconazole-sensitivity and -field efficacy , 2008 .

[174]  J. Fernandez-Cornejo,et al.  Economic and policy issues of U.S. agricultural pesticide use trends. , 2013, Pest management science.

[175]  Ian R. Craig,et al.  Sensitivity of Phakopsora pachyrhizi towards quinone-outside-inhibitors and demethylation-inhibitors, and corresponding resistance mechanisms. , 2014, Pest management science.

[176]  B. Oakley,et al.  Mitosis in Wild-Type and β-Tubulin Mutant Strains ofAspergillus nidulans , 1998 .

[177]  Shengming Liu,et al.  Functional analysis of the β2 -tubulin gene of Fusarium graminearum and the β-tubulin gene of Botrytis cinerea by homologous replacement. , 2013, Pest management science.

[178]  M. Csukai,et al.  Mutagenesis and Functional Studies with Succinate Dehydrogenase Inhibitors in the Wheat Pathogen Mycosphaerella graminicola , 2012, PloS one.

[179]  H. Cools,et al.  Are azole fungicides losing ground against Septoria wheat disease? Resistance mechanisms in Mycosphaerella graminicola. , 2008, Pest management science.

[180]  S. Kaneko,et al.  Identification of three mutant loci conferring carboxin-resistance and development of a novel transformation system in Aspergillus oryzae. , 2009, Fungal genetics and biology : FG & B.

[181]  G. Bergstrom,et al.  Triazole Sensitivity in a Contemporary Population of Fusarium graminearum from New York Wheat and Competitiveness of a Tebuconazole-Resistant Isolate. , 2014, Plant disease.

[182]  T. Michailides,et al.  Sensitivities of Baseline Isolates and Boscalid-Resistant Mutants of Alternaria alternata from Pistachio to Fluopyram, Penthiopyrad, and Fluxapyroxad. , 2014, Plant disease.

[183]  H. Ishii,et al.  Genetic analysis and molecular characterisation of laboratory and field mutants of Botryotinia fuckeliana (Botrytis cinerea) resistant to QoI fungicides. , 2012, Pest management science.

[184]  J. Bohlmann,et al.  The versatility of the fungal cytochrome P450 monooxygenase system is instrumental in xenobiotic detoxification , 2011, Molecular microbiology.

[185]  A. Yahyaoui,et al.  Sensitivity of Mycosphaerella graminicola isolates from Tunisia to epoxiconazole and pyraclostrobin , 2012 .

[186]  C. Xiao,et al.  Molecular characterization of boscalid resistance in field isolates of Botrytis cinerea from apple. , 2011, Phytopathology.

[187]  H. Cools,et al.  Overexpression of the sterol 14α-demethylase gene (MgCYP51) in Mycosphaerella graminicola isolates confers a novel azole fungicide sensitivity phenotype. , 2012, Pest management science.

[188]  T. Anke,et al.  The molecular basis for the natural resistance of the cytochrome bc1 complex from strobilurin-producing basidiomycetes to center Qp inhibitors. , 1996, European journal of biochemistry.

[189]  T. Staub,et al.  Review: Resistance as a concomitant of modern crop protection , 1997 .

[190]  C. Yanofsky,et al.  Cloning and characterization of the gene for beta-tubulin from a benomyl-resistant mutant of Neurospora crassa and its use as a dominant selectable marker , 1986, Molecular and cellular biology.

[191]  H. Cools,et al.  Update on mechanisms of azole resistance in Mycosphaerella graminicola and implications for future control. , 2013, Pest management science.

[192]  P. Leroux,et al.  Evolution of fungicide resistance in the cereal eyespot fungi Tapesia yallundae and Tapesia acuformis in France , 1997 .

[193]  P. E. Russell,et al.  Resistance development to QoI inhibitors in populations of Mycosphaerella graminicola in the UK. , 2005 .

[194]  J. West,et al.  Detection and molecular characterisation of Pyrenopeziza brassicae isolates resistant to methyl benzimidazole carbamates. , 2013, Pest management science.

[195]  J. Peberdy,et al.  Resistance to fungicides in field isolates and laboratory induced mutants of Pseudocercosporella herpotrichoides , 1990 .

[196]  W. Melchers,et al.  Azole resistance in Aspergillus fumigatus: a side-effect of environmental fungicide use? , 2009, The Lancet. Infectious diseases.

[197]  J. Beckerman Detection of Fungicide Resistance , 2013 .

[198]  James K. M. Brown,et al.  Plant-parasite coevolution: bridging the gap between genetics and ecology. , 2011, Annual review of phytopathology.

[199]  G. Schnabel,et al.  The Cytochrome P450 Lanosterol 14α-Demethylase Gene Is a Demethylation Inhibitor Fungicide Resistance Determinant in Monilinia fructicola Field Isolates from Georgia , 2007, Applied and Environmental Microbiology.

[200]  H. Sierotzki,et al.  Mechanisms influencing the evolution of resistance to Qo inhibitor fungicides. , 2002, Pest management science.

[201]  H. Maraite,et al.  Evidence for a heterothallic mating system in Tapesia acuformis using benomyl sensitivity and esterase isoenzyme profiles , 1996 .

[202]  Zahir Eyal,et al.  The Septoria Tritici and Stagonospora Nodorum Blotch Diseases of Wheat , 1999, European Journal of Plant Pathology.

[203]  Fangwei Yu,et al.  Paralogous cyp51 genes in Fusarium graminearum mediate differential sensitivity to sterol demethylation inhibitors. , 2011, Fungal genetics and biology : FG & B.

[204]  Steven L. Kelly,et al.  Mechanism of Binding of Prothioconazole to Mycosphaerella graminicola CYP51 Differs from That of Other Azole Antifungals , 2010, Applied and Environmental Microbiology.

[205]  A. Jones,et al.  The 14alpha-Demethylasse(CYP51A1) Gene is Overexpressed in Venturia inaequalis Strains Resistant to Myclobutanil. , 2001, Phytopathology.

[206]  D. Jones,et al.  Factors affecting diseases of winter wheat in England and Wales, 1989–98 , 2001 .

[207]  B. Spooner,et al.  Tapesia yallundae-the teleomorph of Pseudocercosporella herpotrichoides , 1988 .

[208]  J. Dekker Acquired resistance to fungicides , 1976 .

[209]  J. Beckerman,et al.  In Situ Detection of Benzimidazole Resistance in Field Isolates of Venturia inaequalis in Indiana. , 2010, Plant disease.

[210]  E. Mellado,et al.  Probing the role of point mutations in the cyp51A gene from Aspergillus fumigatus in the model yeast Saccharomyces cerevisiae. , 2011, Medical mycology.

[211]  E. Mellado,et al.  Identification of Two Different 14-α Sterol Demethylase-Related Genes (cyp51A and cyp51B) in Aspergillus fumigatus and Other Aspergillus species , 2001, Journal of Clinical Microbiology.

[212]  H. Deising,et al.  Development of a novel multiplex DNA microarray for Fusarium graminearum and analysis of azole fungicide responses , 2011, BMC Genomics.

[213]  F. Daldal,et al.  A compilation of mutations located in the cytochrome b subunit of the bacterial and mitochondrial bc1 complex. , 1996, Biochimica et biophysica acta.

[214]  D. Backhouse Global distribution of Fusarium graminearum, F. asiaticum and F. boothii from wheat in relation to climate , 2014, European Journal of Plant Pathology.

[215]  P. Esker,et al.  Quantitative review of fungicide efficacy trials for managing soybean rust in Brazil. , 2009 .

[216]  N. Talbot,et al.  A sterol 14α-demethylase is required for conidiation, virulence and for mediating sensitivity to sterol demethylation inhibitors by the rice blast fungus Magnaporthe oryzae. , 2011, Fungal genetics and biology : FG & B.

[217]  Jian-xin Wang,et al.  Mutations in a beta-tubulin confer resistance of Gibberella zeae to benzimidazole fungicides. , 2009, Phytopathology.

[218]  D. Kelly,et al.  Paralog Re-Emergence: A Novel, Historically Contingent Mechanism in the Evolution of Antimicrobial Resistance , 2014, Molecular biology and evolution.

[219]  D. Hollomon,et al.  A critical evaluation of the role of alternative oxidase in the performance of strobilurin and related fungicides acting at the Qo site of complex III. , 2003, Pest management science.

[220]  P. Leroux,et al.  Multiple mechanisms account for resistance to sterol 14α-demethylation inhibitors in field isolates of Mycosphaerella graminicola. , 2011, Pest management science.

[221]  U. Gisi,et al.  Sensitivity profiles of Mycosphaerella graminicola and Phytophthora infestans populations to different classes of fungicides , 1997 .

[222]  H. Cools,et al.  A novel substitution I381V in the sterol 14alpha-demethylase (CYP51) of Mycosphaerella graminicola is differentially selected by azole fungicides. , 2007, Molecular plant pathology.

[223]  M. Shaw,et al.  Effects of fungicide dose and mixtures on selection for triazole resistance in Mycosphaerella graminicola under field conditions , 2006 .

[224]  D. Hollomon,et al.  Effect of carbendazim resistance on trichothecene production and aggressiveness of Fusarium graminearum. , 2009, Molecular plant-microbe interactions : MPMI.

[225]  K. Grossmann,et al.  Metrafenone: studies on the mode of action of a novel cereal powdery mildew fungicide. , 2006, Pest management science.

[226]  V. ter meulen,et al.  How should we tackle the global risks to plant health? , 2014, Trends in plant science.