浙江省西 (甜) 瓜蔓枯病菌对甲基硫菌灵和啶酰菌胺的抗性检测及抗性机制

从浙江省5地采集了112株西 (甜) 瓜蔓枯病菌Stagonosporppsis citrulli,采用区分剂量法检测其对苯并咪唑类杀菌剂甲基硫菌灵 (以下简称Ben) 和琥珀酸脱氢酶抑制剂类 (SDHIs) 杀菌剂啶酰菌胺 (简称Bos) 的抗性。结果显示:112株西 (甜) 瓜蔓枯病菌对Ben和Bos的抗药性频率分别为100%和28.6%,其中对甲基硫菌灵产生高水平抗性 (BenHR) 的菌株达100%,对啶酰菌胺产生低水平抗性 (BosLR) 和高水平抗性 (BosHR) 的菌株分别为18.8%和9.8%。抗药性分子机制研究表明:BenHR菌株中β-tubulin的第198位氨基酸由Glu (E) 突变成了Ala (A);BosHR菌株中Sdh B的第277位氨基酸由His (H) 突变成了Tyr (Y),但BosLR的抗性机制还需进一步研究。研究结果表明,浙江省西 (甜) 瓜蔓枯病菌对苯并咪唑类杀菌剂甲基硫菌灵的抗性已十分严重,尽管大多数菌株对啶酰菌胺仍表现敏感,但在一些地区已存在高水平抗性菌株,应在加强抗药性监测的同时,注意SDHIs类杀菌剂的科学使用。     

吉林省西瓜蔓枯病菌对吡唑醚菌酯的抗性监测及抗性风险评估

为明确吉林省西瓜蔓枯病菌Stagonosporopsis citrulli对吡唑醚菌酯的敏感基线和抗性风险,采用菌丝生长速率法测定吉林省151株西瓜蔓枯病菌菌株对吡唑醚菌酯的敏感性及吡唑醚菌酯对西瓜蔓枯病菌的最低抑制浓度,并监测375株西瓜蔓枯病菌菌株对吡唑醚菌酯的抗性水平。结果表明,供试的151株菌株对吡唑醚菌酯的敏感性频率分布呈连续的单峰曲线,接近正态分布,敏感基线为1.800 μg/mL;吡唑醚菌酯的最低抑制浓度MIC为80.000 μg/mL。抗性水平监测结果显示,375株菌株对吡唑醚菌酯的抗性频率为0。室内药剂驯化获得的抗性突变菌株T-LH3-1和T-XZC5-1在连续转接培养8代后的EC₅₀分别为16.139 μg/mL和31.782 μg/mL,抗性倍数分别为15和51,均为中抗水平,抗药性可以稳定遗传。抗性突变体及其亲本菌株生物学性状结果显示,抗性突变体生物适合度与其亲本菌株差异不显著。交互抗性结果表明,吡唑醚菌酯与醚菌酯之间存在正交互抗性,与戊唑醇、异菌脲、多菌灵和肟菌酯之间不存在交互抗性,可以交替使用以达到延缓抗药性的产生。     

啶酰菌胺不同抗性类型多主棒孢的生物学特性

由多主棒孢Corynespora cassiicola引起的黄瓜棒孢叶斑病是黄瓜上的重要病害。本研究测定了对啶酰菌胺不同抗性类型多主棒孢的生物学特性差异和环境适合度，旨在为探明多主棒孢对啶酰菌胺的抗性变化机制提供生物学研究基础。随机选取不同地区24株对啶酰菌胺具有不同抗性类型的多主棒孢，分析了7种类型抗性突变体对几种不同琥珀酸脱氢酶抑制剂类 (SDHIs) 杀菌剂的交互抗性、不同抗性类型多主棒孢在无药剂选择压力下的抗性遗传稳定性，以及抗性突变体在不同碳源、氮源、温度等环境条件下的生物学特性及适合度。结果表明:除突变类型SdhB-H278Y及SdhB-H278R对啶酰菌胺与氟吡菌酰胺之间存在负交互抗性外，其他突变类型对啶酰菌胺与吡噻菌胺、氟吡菌酰胺及萎锈灵之间均表现为正交互抗性；所有突变类型菌株的抗药性均能稳定遗传；不同突变类型菌株之间致病性存在差异，其中SdhD-D95E突变体的致病力最强；利于所有突变类型菌株生长的碳源是麦芽糖，氮源种类则对突变体的生长影响不显著；各突变类型菌株的最适生长温度范围为25~30 ℃，其中突变体SdhD-D95E在高于30 ℃条件下菌丝生长速率大于其他突变体；耐热性研究中，抗性突变体经65 ℃高温处理45 min后无法存活，同时发现，60 ℃条件下突变体能正常生长，而敏感菌株不能生长；各质量浓度NaCl处理下，SdhD-D95E突变菌株菌丝生长速率快于其他突变菌株，而SdhB-H278Y突变菌株慢于其他突变菌株；葡萄糖对SdhB-H278R突变菌株的生长较为重要。研究表明，对啶酰菌胺不同抗性类型多主棒孢突变菌株的生物学特性及适合度存在差异，SdhD-D95E突变菌株适合度有所提高，表明该突变类型多主棒孢在田间具有较强的竞争力，容易形成优势种群。     

山东省玉米纹枯病菌对噻呋酰胺的敏感性及其抗性突变体的生物学性状

为明确山东省玉米纹枯病菌对噻呋酰胺的敏感性及其抗药性风险,采用菌丝生长速率法分别测定了采自山东泰安、临沂、潍坊、莱芜、日照及青岛6个地区的102株玉米纹枯病菌对噻呋酰胺的敏感性。结果表明:其EC₅₀值分布范围为0.010～0.194 μg/mL,平均EC₅₀ 值为 (0.086 ± 0.004) μg/mL,且敏感性呈连续单峰曲线分布。通过紫外诱导与药剂驯化的方法各获得5株耐药性菌株 (TA3-X2、TA17-X6、LY8-3、QD14-Y7和WF6-A2) 和1株抗性突变体 (QD2-Y4),其抗性水平在6.46～20.08倍之间,突变频率分别为0.87%和0.52%。对抗性突变体生物学性状的研究表明,紫外诱导获得的5株耐药性菌株其耐药性不能稳定遗传,而经药剂驯化获得的1株抗性突变体QD2-Y4的抗药性可稳定遗传;耐药性菌株TA3-X2的菌丝生长速率高于亲本菌株,其余菌株与亲本菌株差异不明显;5株耐药性菌株和1株抗性突变体的菌丝干重和菌核干重均低于亲本菌株;TA3-X2、WF6-A2及QD2-Y4的致病力低于亲本菌株,TA17-X6、LY8-3及QD14-Y7的致病力与亲本菌株无明显差异。交互抗性测定表明,噻呋酰胺抗性突变体与戊唑醇、丙环唑、咯菌腈、井冈霉素、苯醚甲环唑和多菌灵之间均无交互抗性,与啶酰菌胺和氟唑菌苯胺之间则存在交互抗性。研究表明,山东省6个地区的玉米纹枯病菌对噻呋酰胺比较敏感,推测噻呋酰胺可作为防治玉米纹枯病的理想候选药剂。     

樱桃褐腐病菌对啶酰菌胺的敏感性及其对4种琥珀酸脱氢酶抑制剂的交互抗性

采用菌丝生长速率法测定了樱桃褐腐病菌Monilinia fructicola对啶酰菌胺的敏感性,同时研究了不同敏感性菌株的生物学性状,探究了琥珀酸脱氢酶B亚基的氨基酸突变与其对啶酰菌胺产生抗性的相关性,并分析了樱桃褐腐病菌对啶酰菌胺与其他3种琥珀酸脱氢酶抑制剂(SDHIs)氯苯醚酰胺、氟唑菌苯胺和氟吡菌酰胺之间的交互抗...     

抗啶酰菌胺蔬菜灰霉病菌突变体的诱导及其生物学性状

为评价蔬菜灰霉病菌对啶酰菌胺的抗性风险,采用紫外线照射和药剂驯化相结合的诱导方法,对蔬菜灰霉病菌D9、L17、J9亲本敏感菌株进行了室内啶酰菌胺抗性诱导,并采用菌落直径法比较了抗、感菌株在生物学性状方面的差异。结果显示,共获得不同抗性水平的突变体23株,其中突变体D9-4的抗性倍数为253.76倍,达高抗水平。突变体菌丝生长速率较敏感菌株有所下降,抗、感菌株的适宜生长温度均为25 ℃,敏感菌株最适pH为5,突变体最适pH为7,突变体适应酸碱度和温度变化的能力较敏感菌株差;低抗突变体产孢量和孢子萌发率与敏感菌株无显著差异,但高抗突变体产孢量和孢子萌发率较敏感菌株显著降低;在无药条件下,敏感菌株致病力强于抗性突变体,而在药剂胁迫下,突变体表现出一定的耐药性,致病力强于敏感菌株;6株突变体无药继代培养15代后,突变体抗性稳定遗传。此外,蔬菜灰霉病菌对啶酰菌胺与其它5种杀菌剂之间没有交互抗性。研究表明,蔬菜灰霉病菌抗啶酰菌胺突变体适合度较低,但在药剂选择压力下容易形成啶酰菌胺抗性群体,在生产中需引起注意。     

番茄灰霉病菌对咯菌腈的抗性诱变及抗药突变体的生物学特性

为评估番茄灰霉病菌Botrytis cinerea对咯菌腈的抗性风险,就室内经紫外照射获得抗药突变体的方法及抗性突变体的生物学性状进行了研究。结果表明:番茄灰霉病菌分生孢子的紫外照射亚致死时间为90~120 s;经亚致死时间紫外照射后,4个亲本菌株中有2个菌株共产生了6个抗咯菌腈的突变体,其EC50值是亲本菌株的310倍以上,抗性突变频率为3.13×10-7;经紫外照射诱变获得的所有抗性突变体在菌丝生长速率、产孢量、产菌核能力及其在番茄果实上的致病性方面均比其亲本菌株明显降低。相关分析显示,所得抗咯菌腈突变体对氟啶胺、啶菌唑、啶酰菌胺和嘧霉胺无交互抗性。表明番茄灰霉病菌对咯菌腈的抗药性风险较低。     

贵州省烟草赤星病菌对啶酰菌胺的敏感性基线

对2014—2015年采自贵州省田间未使用过琥珀酸脱氢酶抑制剂 (SDHIs) 的烟草赤星病病样进行了病原菌分离和鉴定,并采用菌丝生长速率法建立了烟草赤星病菌群体对啶酰菌胺的敏感性基线。结果表明:虽然烟草赤星病菌不同菌株在菌落、孢子形态和致病性上有较大差异,但均属于交链格孢Alternaria alternata。啶酰菌胺抑制赤星病菌群体 (n = 102) 菌丝生长的EC₅₀值在0.186~5.818 μg/mL之间,平均EC₅₀值为 (2.157 ± 1.112) μg/mL,不同敏感性菌株频率分布为连续的单峰曲线,该平均EC₅₀值可以作为烟草赤星病菌对啶酰菌胺的敏感性基线。进一步研究表明,贵州省烟草赤星病菌的Sdh B基因在第209位、277位和224位等密码子存在核苷酸多样性,但未发现这种多样性与烟草赤星病菌对啶酰菌胺的敏感性之间存在关联。     

白术两种主要土传病害的分离、鉴定及杀菌剂的室内活性筛选

立枯病和白绢病是白术生产种植中的两种主要土传病害,在苗期和生长期都有发生,危害严重。本研究从浙江省磐安县采集具有典型病症的白术植株,对病原进行了分离、纯化和致病性测定。综合形态学特征及rDNA-ITS序列分析表明,白术上的病害是由立枯丝核菌Rhizoctonia solani Kühn引起的立枯病和由齐整小核菌Sclerotium rolfsii Sacc.引起的白绢病。室内毒力测定结果表明,10种供试杀菌剂对立枯病菌的毒力从大到小为:噻呋酰胺咯菌腈戊唑醇四氟醚唑吡唑醚菌酯丙硫菌唑嘧菌酯啶酰菌胺苯醚甲环唑咪鲜胺;对白绢病菌的室内毒力从大到小为:吡唑醚菌酯噻呋酰胺嘧菌酯戊唑醇咯菌腈苯醚甲环唑丙硫菌唑啶酰菌胺四氟醚唑咪鲜胺,其中噻呋酰胺对两种病菌都具有很高的活性,EC_(50)分别为0.06和0.03mg/L,可用于两种病害的防治。     

美澳型核果褐腐病菌对甲基硫菌灵和啶酰菌胺的敏感性

采用传统检测方法测定了我国北方地区美澳型核果褐腐病菌对常用杀菌剂甲基硫菌灵和新药剂啶酰菌胺的敏感性,并采用12条ISSR引物对不同抗性水平的菌株进行了分析.结果显示:74株菌株中检测出5株对甲基硫菌灵表现高抗、2株表现低抗的菌株;我国抗性菌株的突变位点与美国加州抗性菌株的突变位点相同;啶酰菌胺对来自我国和美国、新西兰、法国的45株美澳型核果褐腐病菌的EC50分布频率呈单峰曲线,EC50在0.059～0.320 μg/mL之间,平均值为0.174±0.064μg/mL;基于ISSR分析得到的UPGMA聚类组与菌株的来源地及对甲基硫菌灵的抗性无相关性.     

Characterization of mutations in the iron-sulphur subunit of succinate dehydrogenase correlating with Boscalid resistance in Alternaria alternata from California pistachio

Thirty-eight isolates of Alternaria alternata from pistachio orchards with a history of Pristine (pyraclostrobin + boscalid) applications and displaying high levels of resistance to boscalid fungicide (mean EC(50) values >500 microg/ml) were identified following mycelial growth tests. A cross-resistance study revealed that the same isolates were also resistant to carboxin, a known inhibitor of succinate dehydrogenase (Sdh). To determine the genetic basis of boscalid resistance in A. alternata the entire iron sulphur gene (AaSdhB) was isolated from a fungicide-sensitive isolate. The deduced amino-acid sequence showed high similarity with iron sulphur proteins (Ip) from other organisms. Comparison of AaSdhB full sequences from sensitive and resistant isolates revealed that a highly conserved histidine residue (codon CAC in sensitive isolates) was converted to either tyrosine (codon TAC, type I mutants) or arginine (codon CGC, type II mutants) at position 277. In other fungal species this residue is involved in carboxamide resistance. In this study, 10 and 5 mutants were of type I and type II respectively, while 23 other resistant isolates (type III mutants) had no mutation in the histidine codon. The point mutation detected in type I mutants was used to design a pair of allele-specific polymerase chain reaction (PCR) primers to facilitate rapid detection. A PCR-restriction fragment length polymorphism (RFLP) assay in which amplified gene fragments were digested with AciI was successfully employed for the diagnosis of type II mutants. The relevance of these modifications in A. alternata AaSdhB sequence in conferring boscalid resistance is discussed.     

Resistance profiles of Botrytis cinerea populations to several fungicide classes on greenhouse tomato and strawberry in Lebanon

Tomato and strawberry are the most important protected crops in Lebanon and are seriously affected by grey mould disease, caused by Botrytis cinerea. In the present study, the fungicide sensitivity assays revealed medium to high frequencies of B. cinerea isolates resistant to benzimidazoles, dicarboximides, and anilinopyrimidines on tomato and strawberry. Fludioxonil- and boscalid-resistant mutants were uncommonly found at generally low frequency on both crops. Resistance to fenhexamid was detected in only one site on tomato but in most sites on strawberry with high frequencies, and the occurrence of resistance to QoI fungicides was ascertained on both crops. The majority of the tested isolates (>90%) exhibited multiple fungicide resistance, and isolates resistant to the seven antibotrydial fungicide classes were detected on strawberry in three locations. A high level of resistance was shown by B. cinerea mutants resistant to boscalid, fenhexamid, and QoI fungicides, while two levels of moderate and high resistance to anilinopyrimidines were identified. Genetic analysis revealed point mutations in the target genes commonly associated with resistance in B. cinerea isolates, with all mutants resistant to dicarboximides, fenhexamid, boscalid, and QoI fungicides carrying single-nucleotide polymorphims in BcOS1 (I365S/N, Q369P, and N373S), Erg27 (F412V/I), SdhB (H272R/Y), and cytb (G143A) genes, respectively. The general incorrect use of fungicides has caused the development and spread of fungicide resistance as a widespread phenomenon on protected tomato and strawberry in Lebanon. The implementation of appropriate antiresistance strategies is highly recommended.     

Characterization of mutations in the membrane-anchored subunits AaSDHC and AaSDHD of succinate dehydrogenase from Alternaria alternata isolates conferring field resistance to the fungicide boscalid

The molecular basis of resistance to the fungicide boscalid in 25 Alternaria alternata field mutants exhibiting resistance to boscalid and previously tested negative for AaSDHB mutations conferring boscalid resistance was investigated by cloning and sequencing the A. alternata SDHC ( AaSDHC ) and SDHD ( AaSDHD ) genes from a boscalid-sensitive isolate. The SDHB and SDHC/SDHD genes encode the iron sulphur and two membrane-anchored subunits of succinate:ubiquinone oxidoreductase (SQR) that constitute the boscalid fungicide molecular targets. The deduced amino acid sequences exhibited low similarities with SDHC and SDHD peptides from other organisms, but residues essential to form the ubiquinone binding site or important in SQR assembly were particularly conserved. Sequence comparisons of the AaSDHC and AaSDHD genes between resistant mutants and wild-type isolates revealed that two highly conserved histidine residues implicated in the heme b ligation and located at codon 134 in AaSDHC (22 mutants) and codon 133 in AaSDHD (two mutants) were replaced by arginine residues (H134R and H133R). In another mutant, a substitution of an aspartate by a glutamic acid occurred at amino acid position 123 (D123E) in AaSDHD. Additional tests revealed that mycelial growth of boscalid-resistant isolates was reduced when isolates were subjected to oxidative stress. The identified mutations were confirmed using PCR-RFLP assays. This is apparently the first report of mutations located in the heme b ligands of the cytochrome II gene associated with carboxamide resistance.     

浙江省果蔬灰葡萄孢对啶酰菌胺的抗性

以2004—2006年从浙江、江苏等地采集的灰葡萄孢对啶酰菌胺的敏感性基线[EC₅₀ = (1.07 ± 0.11) mg/L]为依据,采用菌丝生长速率法连续监测了浙江省果蔬灰葡萄孢群体对啶酰菌胺的敏感性变化。结果表明:浙江省果蔬灰葡萄孢对啶酰菌胺的抗性发展迅速,2012—2013年和2017—2018年的平均EC₅₀值分别为 (5.23 ± 7.79) 和 (24.30 ± 49.33) mg/L。其中,2012—2013年的抗药性菌株频率为15.3%,且均为低水平抗性 (LR) 菌株;而2017—2018年的抗药性频率上升至53.2%,并出现了7.5%的中等水平抗性 (MR) 菌株和1.3%的高水平抗性 (HR) 菌株。啶酰菌胺抗性菌株的菌丝生长速率、产孢量、产菌核数和致病力与敏感菌株相比均无显著差异。抗药性分子机制研究表明:啶酰菌胺抗性菌株的琥珀酸脱氢酶B亚基 (SDH B) 均发生了点突变,共包括H272R、P225F和N230I 3种类型,其中H272R型突变占88.5%;其SDH A和SDH D均未发生点突变;而SDH C的突变 (G85A + I93V + M158V + V168I) 与对药剂敏感性之间无明显联系。     

Biological and molecular characterization of field isolates of <Emphasis Type="Italic">Alternaria alternata</Emphasis> with single or double resistance to respiratory complex II and III inhibitors

Field isolates of Alternaria alternata collected from tomato processors were characterized for sensitivity to respiration inhibitors using in vitro mycelial growth assays. Pyraclostrobin (QoI), boscalid, fluopyram and isopyrazam (SDHIs) mean EC₅₀ values were 0.32, 1.43, 2.21, and 3.53 μg/ml respectively. Of the 42 isolates, 36 were sensitive to all respiration inhibiting fungicides tested whereas three isolates were less sensitive to boscalid, one to pyraclostrobin and two were simultaneously resistant to both inhibitors and isopyrazam. Correlation analysis between fungicide sensitivities revealed a positive cross-resistance between pyraclostrobin and tebuconazole, and between cyprodinil and mancozeb. There was no cross-resistance between QoIs, SHDIs or any other mode of action. Sequencing of the QoI and SDHI targets revealed the G143A cytochrome b resistance mutation in all pyraclostrobin-resistant isolates while analysis of the succinate dehydrogenase coding gene revealed point mutations in two of three of the gene subunits analyzed in boscalid-resistant isolates. Specifically, two isolates carried the H277Y and three the H133Q resistance mutations located in the sdhB and sdhD subunits of the respiration complex II, respectively. Isolates bearing the H277Y mutation also carried the G143A cytochrome b resistance mutation. Boscalid and pyraclostrobin-resistant isolates exhibited greater pathogenicity and sporulation compared to sensitive isolates, respectively. Isolates with cross-resistance exhibited greater pathogenicity and sporulation but slower mycelial growth compared to sensitive isolates. This is the first report of field isolates of A. alternata with single or double resistance to QoIs and SDHIs in Greece and should be considered in planning and implementing effective anti-resistance strategies.     

Sensitivity of Botrytis cinerea from vegetable greenhouses to boscalid

Between 2004 and 2006, 228 isolates of Botrytis cinerea from two regions in China were characterized for baseline sensitivity to boscalid, a new active ingredient that interferes with succinate ubiquinone reductase in the electron transport chain. The isolates showed similar sensitivity in different years and regions. Baseline sensitivities were distributed as unimodal curves with mean EC₅₀ values of 1·07 (± 0·11) and 0·42 (± 0·05) mg L⁻¹ for inhibition of mycelial growth and conidial germination, respectively. Laboratory studies were conducted to evaluate the risk of development of resistance to boscalid. Boscalid-resistant mutants were obtained by UV-treatment at lower frequencies and with smaller resistance factors than pyrimethanil-resistant mutants. All boscalid-resistant mutants were also significantly more sensitive to Qo inhibitors than their wild-type parents and showed reduced sporulation in vitro and pathogenicity on aubergine leaves. The results suggested that the risk of resistance developing for boscalid was lower than for pyrimethanil. However, as B. cinerea is a high-risk pathogen, appropriate precautions against resistance development should be taken. Synergism between the activity of boscalid and that of kresoxim-methyl was observed.     

抗苯醚菌酯(ZJ0712)小麦白粉病菌突变体的诱导及其生物学特性

经过药剂驯化10代后,从对苯醚菌酯(ZJ0712)敏感的小麦白粉病菌Blumeria graminis f.sp.tritici 3个菌株中获得3个抗药突变体,突变体的抗性指数均大于80,且其抗性能够通过无性繁殖稳定遗传。室内接种试验发现,突变体的致病力与亲本菌株无明显差异。细胞色素b (cyt b)基因片段序列分析发现,突变体cyt b基因的第143位密码子均由敏感菌株的GGT(丙氨酸)突变成了GCT(甘氨酸)。研究结果表明,小麦白粉病菌对苯醚菌酯存在较高的抗性风险,该药剂在使用过程中需注意采取相应的抗性治理措施,以延缓抗性发生。     

小麦纹枯病菌对噻呋酰胺的敏感性及抗药性突变体的主要生物学性状

采用菌丝生长速率法,分别测定了采自河北、山东和河南未使用过酰胺类药剂地区的166株小麦纹枯病菌Rhizotonia cerealis Van der Hoeven对噻呋酰胺的敏感性,并对小麦纹枯病菌抗噻呋酰胺突变体的诱导方法及突变体的主要生物学性状进行了研究。结果表明:噻呋酰胺对166株小麦纹枯病菌的平均EC50值为(0.072±0.022)μg/mL,菌株频率呈连续单峰曲线分布,未发现敏感性明显下降的亚群体,因此可将该EC50值作为小麦纹枯病菌对噻呋酰胺的敏感基线。通过紫外诱导加菌落角变的方法获得了5株抗噻呋酰胺突变体,其抗药性水平介于5.5~18.9倍之间;药剂驯化未能获得抗药性突变体。突变体在PSA平板上的生长速率与亲本菌株无显著性差异,但其菌丝干重、菌核形成数量和菌核干重均明显低于亲本菌株;除突变体HD7-7U对石新828的致病力未发生明显变化外,其他突变体对石新828和良星99的致病力均明显减弱;5株突变体的抗药性均不能稳定遗传;噻呋酰胺与戊唑醇、井冈霉素、咯菌腈、三唑酮、丙环唑和多菌灵之间无交互抗性关系。     

Impact of fludioxonil resistance on fitness and cross-resistance profiles of Altrenaria solani laboratory mutants

Sensitivity and inherent resistance risk of Alternaria solani to fludioxonil, cross-resistance profiles and the potential implications of resistance mutations on fitness parameters were investigated. Fludioxonil was highly effective against a wild type A. solani field strain both in vitro (EC₅₀?=?0.05 μg/mL) and in preventive applications on artificially inoculated tomato fruit. Mutants with low [Resistance factor (Rf): 15 based on EC₅₀], medium (Rf: 150–300) and high (Rf: > 1000) levels of phenylpyrrole resistance were isolated from the wild type strain at high frequencies following mutagenesis with UV irradiation and selection on fludioxonil containing medium. Resistant isolates retained their resistance levels even after 9 subcultures on fungicide-free growth medium while they could express their resistant phenotypes in planta. Investigation of cross-resistance relationships showed that fludioxonil resistance mutations also reduce the sensitivity of mutant strains to the aromatic hydrocarbon fungicide quintozene as well as the dicarboximides iprodione and vinclozolin. No cross-resistance was observed between fludioxonil and fungicides with different modes of action such as the sterol biosynthesis inhibitors (DMIs) imazalil and flusilazole and the carboxamide boscalid. All fludioxonil resistant isolates were more sensitive to the anilinopyrimidine pyrimethanil, while only two isolates were less sensitive to the QoI pyraclostrobin compared to the wild-type strain. Study of fitness determining parameters showed that resistance mutation(s) had no adverse effects on mycelial growth, conidial germination and sensitivity to osmotic stress while they had a pleiotropic effect on virulence and conidia production in resistant mutants. Results of the present study indicate that fludioxonil is a highly effective fungicide against A. solani, while the risk of resistance development to this fungicide is considered to be medium making fludioxonil an ideal alternative to high risk fungicides such as boscalid and pyraclostrobin whose performance against early blight has already been compromised by resistance development.