Transfer of the β‐tubulin gene of Botrytis cinerea with resistance to carbendazim into Fusarium graminearum

BACKGROUND: Resistance to carbendazim and other benzimidazole fungicides in Botrytis cinerea (Pers. ex Fr.) and most other fungi is usually conferred by mutation(s) in a single chromosomal β‐tubulin gene, often with several allelic mutations. In Fusarium graminearum Schwade, however, carbendazim resistance is not associated with a mutation in the corresponding β‐tubulin gene. RESULTS: The β‐tubulin gene conferring carbendazim resistance in B. cinerea was cloned and connected with two homologous arms of the β‐tubulin gene of F. graminearum by using a double‐joint polymerase chain reaction (PCR). This fragment was transferred into F. graminearum via homologous double crossover at the site where the β‐tubulin gene of F. graminearum is normally located (the β‐tubulin gene of F. graminearum had been deleted). The transformants were confirmed and tested for their sensitivity to carbendazim. CONCLUSION: The β‐tubulin gene conferring carbendazim resistance in B. cinerea could not express this resistance in F. graminearum, as transformants were still very sensitive to carbendazim. Copyright © 2010 Society of Chemical Industry     

Further studies in increased sensitivity to N-phenylanilines in benzimidazole-resistant strains of Botrytis cinerea and Venturia nashicola

The ring-substituted N-phenylanilines, N-(3-chlorophenyl)aniline (MC-1) and N-(3,5-dichlorophenyl)aniline (MC-2) were tested for their antifungal activity against Botrytis cinerea Pers. ex Fr. and Venturia nashicola Tanaka et Yamamoto. In both fungi, increased sensitivity to MC-1 and MC-2 was clearly observed in ‘highly carbendazim-resistant, diethofencarb-sensitive’ (HR, S) phenotypes. Sensitivity was low in ‘carbendazim-sensitive, diethofencarb-resistant’ (S, R) and ‘intermediately carbendazim-resistant, diethofencarb-resistant’ (IR, R) strains. On cucumber cotyledons, other strains of B. cinerea, possessing the phenotype ‘highly carbendazim-resistant, diethofencarb-resistant’ (HR, R) were not controlled by either MC-1 or MC-2. Response to MC-2 was also examined using random ascospore progenies from V. nashicola crosses. In these progenies, high-level carbendazim resistance and MC-2 sensitivity always segregated together. Sensitivity to MC-2 is controlled by a single gene which is either identical to or very closely linked to one conferring high-level resistance to carbendazim.     

应用电导仪测定番茄灰霉病菌对多菌灵抗药性的初步研究

1.0 mg/L和5.0 mg/L多菌灵盐酸盐溶液处理灰霉病菌多菌灵敏感菌株和抗性菌丝体,用电导仪测定溶液电导率的动态变化,结果表明:在敏感菌株存在时,多菌灵盐酸盐溶液2 h后的电导率显著降低,而抗性菌株存在的溶液中电导率则未见下降。     

Differential binding of a N-phenylformamidoxime compound in cell-free extracts of benzimidazole-resistant and-sensitive isolates of Venturia nashicola,Botrytis cinerea and Gibberella fujikuroi

Venturia nashicola isolates with a high level of resistance to carbendazim showed either increased sensitivity or were resistant to theN-phenylformamidoxime compoundN-(3,5-dichloro-4-propynyloyphenyl)-N′-methoxyformamidine (DCPF). Isolates with an intermediate or low level of carbendazim resistance were resistant to DCPF. Increased sensitivity to DCPF was also associated with a high level of carbendazim resistance inBotrytis cinerea but not with a moderate resistance level. Increased sensitivity to DCPF was not observed in carbendazim resistant isolates ofGibberella fujikuroi.Binding of [¹⁴C]DCPF in cell-free mycelial extracts of the highly carbendazim-resistant and DCPF-sensitive isolate ofV. nashicola was higher than in those of DCPF-resistant isolates that were either highly-resistant, intermediately resistant, or weakly resistant to carbendazim or were sensitive to carbendazim. [¹⁴C]carbendazim binding in extracts of highly carbendazim-resistant isolates ofB. cinerea was lower than that in extracts of sensitive isolates, whereas [¹⁴C]DCFP binding was higher. A decreased [¹⁴C]carbendazim binding was also observed in extracts of carbendazim-resistant isolates ofG. fujikuroi, binding of [¹⁴C]DCPF, however, was similar in extracts of both carbendazim-resistant and sensitive isolates.     

Cross-resistance relationships between benzimidazole fungicides and diethofencarb in Botrytis cinerea and their genetical basis in Ustilago maydis

After nitrosoguanidine- or UV-mutagenesis, three different benzimidazole-resistant phenotypes were isolated on media containing benomyl or a mixture of carbendazim and diethofencarb from wild-type strains of Botrytis cinerea Pers. ex Fr. and Ustilago maydis (D.C.) Corda. Mutants of B. cinerea with moderate (MBr) or low (LBr) resistance to benzimidazoles and high resistance to diethofencarb (Dr) were isolated from the fungicide-mixture-containing medium in low frequency (7–1 × 10^?8). Only benzimidazole-resistant strains highly sensitive to diethofencarb (HBrDs) were identified on benomyl-containing medium at a frequency of 6.6 × 10^?6. Fitness-determining characteristics such as sporulation, germination and germ-tube elongation, were found to be reduced significantly in the mutants of B. cinerea that were resistant to both benzimidazoles and diethofencarb. However, pathogenicity of a MBrDr mutant strain on cucumber seedlings was equal to that of the wild type and a carbendazim + diethofencarb mixture was found to control grey mould caused by the wild type, but was not effective when the plant cotyledons were infected by the mutant strain. Three benzimidazole-resistant phenotypes (HBrDs, HBrDr, MBrDr) were isolated easily in U. maydis from a benomyl-containing medium. In contrast with B. cinerea, only one-tenth of the benzimidazole-resistant strains were sensitive to diethofencarb. Genetic analysis of benzimidazole resistance in U. maydis showed that the three benzimidazole-resistant phenotypes were due to three allelic mutations in a single gene and one of them was responsible for the negative cross-resistance between benzimidazoles and diethofencarb.     

禾谷镰孢菌对氟唑菌酰羟胺敏感性基线的建立及药剂田间防效

为明确琥珀酸脱氢酶抑制剂类新型吡啶酰胺杀菌剂氟唑菌酰羟胺在中国小麦赤霉病防治中的应用潜力,分别采用菌丝生长速率法和孢子萌发法,测定了氟唑菌酰羟胺对湖北省6个地区106株禾谷镰孢菌的室内毒力、田间防效及其与多菌灵和氰烯菌酯的交互抗性。结果显示:氟唑菌酰羟胺对106株禾谷镰孢菌菌丝生长的EC₅₀值为 (0.018 0 ± 0.209 0) mg/L,平均值为 (0.072 8 ± 0.025 9) mg/L;对分生孢子萌发的EC₅₀值为 (0.052 7 ± 0.473 2) mg/L,平均值为 (0.176 0± 0.059 6) mg/mL;且其EC₅₀值频率分布均呈单峰曲线,因此可分别将菌丝生长和孢子萌发的平均EC₅₀值作为禾谷镰孢菌对氟唑菌酰羟胺的敏感性基线。初步的交互抗性测定结果表明,抗多菌灵或氰烯菌酯的菌株对氟唑菌酰羟胺均未表现出抗性。田间试验显示,氟唑菌酰羟胺有效剂量200 g/hm2处理的防效 (超过90.0%) 显著高于对照药剂氰烯菌酯600 g/hm2的防效 (78.0%),与空白对照相比增产效果在127%~135%之间。经氟唑菌酰羟胺处理后,小麦籽粒中由禾谷镰孢菌产生的毒素脱氧雪腐镰刀菌烯醇 (DON) 的含量比空白对照降低了55.09%。研究表明,氟唑菌酰羟胺对禾谷镰孢菌呈现出较高的室内活性且田间防效优越,同时还能降低小麦籽粒中DON毒素的含量及提高小麦产量,因此可作为生产中防治小麦赤霉病的替代或后备药剂,同时也可考虑用作为禾谷镰孢菌对多菌灵抗性治理的替代药剂。     

小麦赤霉病菌多菌灵抗性群体的扩散路径分析——基于致病菌种类及所产毒素化学型鉴定和抗药性检出的时序性

通过对江苏、安徽、山东、河南、湖北、河北和四川7省小麦赤霉病菌对多菌灵抗性及敏感菌株Fusarium asiaticum和F.graminearum的鉴定、所产生毒素的化学型及多菌灵抗性菌株检出时序性的分析,初步推测了小麦赤霉病菌对多菌灵抗性群体在中国麦区的扩散路径。结果表明:江淮流域的江苏、安徽、湖北3省和四川省小麦赤霉病菌对多菌灵的抗性或敏感菌株优势群体均是F.asiaticum,而黄淮流域的山东、河南2省及河北省小麦赤霉病菌对多菌灵的敏感菌株优势群体为F.graminearum,抗性菌株优势群体则为F.asiaticum。江苏、安徽、山东和河南抗多菌灵菌株F.asiaticum产生毒素的化学型为3-AcDON和NIV,并以3-AcDON为主。江苏省连续使用多菌灵防治小麦赤霉病长达20多年后才检测到田间抗性菌株,而近年来检测到田间抗性菌株的山东、河南2省用多菌灵防治赤霉病的历史较短,且为偶尔使用,药剂的选择压力相对较小,因此推测山东和河南麦区出现的小麦赤霉病菌抗多菌灵菌株可能是通过种子调运及联合收割机跨区作业等方式从抗药性发生较早的江淮麦区流入的。     

产香真菌GS-1菌株鉴定及其挥发性物质对番茄灰霉病的生防效果

为鉴定产香真菌GS-1菌株物种分类和评价其挥发性物质对番茄灰霉病的生防效果,通过形态学和ITS(internal transcribed spacer)序列分析对菌株GS-1进行物种鉴定,以对扣法测定其挥发物对番茄灰霉病菌Botrytis cinerea、水稻胡麻斑病菌Bipolaris oryzae、小麦赤霉病菌Fusarium graminearum、白菜黑斑病菌Alternaria brassicae和莴苣菌核病菌Sclerotinia sclerotiorum共5种植物病原真菌的熏蒸抑制活性,以果实针刺法测定挥发物对番茄灰霉病的抑制作用,并观察对其菌丝生长的影响。结果显示,菌株GS-1为普通瘤座孢Tubercularia vulgaris;其挥发物对5种植物病原真菌均有明显的熏蒸抑制作用,72 h的抑制率为58.05%~75.81%;其挥发物可造成番茄灰霉病菌菌丝畸形、分枝增多、原生质外渗等,对番茄果实灰霉病的防效72 h后达53.62%。表明产香真菌普通瘤座孢GS-1菌株挥发物对番茄灰霉病有良好的生防效果。     

The effect of benomyl and carbendazim on mitosis in hyphae of Botrytis cinerea Pers. ex Fr. and roots of Allium cepa L

The effects of benomyl [methyl 1 (butylcarbamoyl)-benzimidazol-2-ylcarbamate] and carbendazim (methyl benzimidazol-2-ylcarbamate) on mitosis in hyphae of Botrytis cinerea and root tips of onion (Allium cepa) were studied. The effect of the fungicides on root tips was compared with the effect produced by griseofulvin.Benomyl or carbendazim had a rapid effect on dividing hyphal nuclei. The effects of the fungicides were apparent after 5 min, and the normal stages of division were not seen clearly in any of the treated material. Chromosomes became visible at prophase-metaphase, but they did not separate completely. At the end of division, the chromatin became stretched into long threads, daughter nuclei did not separate completely, and chromatin was often present as irregular shaped masses.Benomyl and carbendazim induced abnormalities in cell division in about 3% of onion root tip cells. After treatment for 6 h, formation of the cell plate was inhibited and cells were seen with two nuclei; other aberrations produced were lagging chromosomes and anaphase bridges. After 3 days chromosomes sometimes occurred as amorphous clumps, and nuclei were found with more than two nucleoli. Griseofulvin produced similar effects in onion root tip cells, but the chromosome bridges persisted through to telophase.The results show that benomyl and carbendazim can interfere with mitosis in cells in green plants as well as in hyphae of fungi.     

玉米内生菌L10的分离、鉴定及拮抗活性

为获得对玉米茎腐病主要病原菌禾谷镰孢Fusarium graminearum有明显拮抗作用的玉米内生菌,采用平板对峙法从成熟健康玉米茎秆中筛选禾谷镰孢拮抗菌株,并分析其抗菌谱;通过形态特征、生理生化特性及16S rDNA序列分析进行菌种鉴定;利用盆栽生防试验检测其对玉米茎腐病的防治效果。结果表明,共分离获得了164株玉米内生细菌菌株,其中L10菌株对禾谷镰孢具有较好的抑制效果,抑菌圈半径达1.68 cm;该菌对玉米大斑病菌Setosphaeria turcica、层出镰孢F. proliferatum、禾谷镰孢F. graminearum、拟轮枝镰孢F. verticilliodes、玉米弯孢叶斑病菌Curvularia lunata、玉米小斑病菌Bipolaris maydis、立枯丝核菌Rhizoctonia solani、茄链格孢Alternaria solani共8种植物病原菌均有拮抗作用,尤其对禾谷镰孢抑制效果最佳;结合形态特征、生理生化性质及16S rDNA序列分析,将L10菌株鉴定为多粘类芽胞杆菌Paenibacillus polymyxa。L10菌株脂肽类物质对禾谷镰孢菌具有较好的抑制活性,且盆栽生防试验结果显示该菌株对玉米茎腐病具有一定的防治效果。表明菌株L10对玉米镰孢茎腐病的防治具有一定潜力。     

Effect of Phenylpyrrole-resistance Mutations on Ecological Fitness of <Emphasis Type="Italic">Botrytis cinerea</Emphasis> and their Genetical Basis in <Emphasis Type="Italic">Ustilago maydis</Emphasis>

Mutants of Botrytis cinerea and Ustilago maydis highly resistant to fludioxonil were isolated at a high frequency, after nitrosoguanidine or UV mutagenesis, respectively, and selection on media containing fludioxonil. Tests on the response of mutant strains to high osmotic pressure resulted in the identification of two fludioxonil-resistant phenotypes (FLD_osm/s and FLD_osm/r), regarding the sensitivity to high osmolarity. Approximately 95% of fludioxonil-resistant mutants were found to be more sensitive to high osmotic pressure than the wild-type parent strains. Genetic analysis of phenylpyrrole-resistance in the phytopathogenic basidiomycete U. maydis, showed that fludioxonil-resistance was coded by three unlinked chromosomal loci (U/fld-1, U/fld-2 and U/fld-3), from which only the U/fld-1 mutation coded an osmotic sensitivity similar to that of the wild-types. Cross-resistance studies with fungicides from other chemical groups showed that the mutations for resistance to phenylpyrroles affect the sensitivity of mutant strains to the aromatic hydrocarbon and dicarboximide fungicides, but not to the benzimidazoles, anilinopyrimidines, phenylpyridinamines, hydroxyanilides or the sterol biosynthesis inhibiting fungicides. A study of fitness parameters in the wild-type and fludioxonil-resistant mutants of B. cinerea, showed that all osmotic sensitive (B/FLD_osm/s) isolates had significant reductions in the characteristics determining saprophytic fitness such as mycelial growth, sporulation, conidial germination and sclerotial production. Contrary to that, with the exception of mycelial growth, the fitness parameters were unaffected or only slightly affected in most of the osmotic resistant (B/FLD_osm/r) isolates. Tests on cucumber seedlings showed that the osmotic-sensitive strains were significantly less pathogenic compared with the wild-type and B/FLD_osm/r strains of B. cinerea. Preventative applications of the commercial products Saphire 50 WP (fludioxonil) and Rovral 50 WP (iprodione) were effective against lesion development on cotyledons by the wild-type and the mutant strains of B. cinerea that were resistant to the anilinopyrimidine cyprodinil (B/CPL-27) and to the hydroxyanilide fenhexamid (B/FNH-21), but ineffective, even at high concentrations, against disease caused by the fludioxonil-resistant isolates (B/FLD) and a mutant strain resistant to the dicarboximide iprodione (B/IPR-1). Experiments on the stability of the fludioxonil-resistant phenotype showed a reduction of resistance, mainly in osmotic-sensitive isolates, when the mutants were grown on inhibitor-free medium. A rapid recovery of the high resistance was observed after mutants were returned to the selection medium. Studies on the competitive ability of mutant isolates against the wild-type parent strain of B. cinerea, by applications of a mixed conidial population, showed that, in vitro, all mutants were less competitive than the wild-type strain. However, the competitive ability of osmotic-resistant mutants was higher than the osmotic-sensitive ones. Furthermore, competition tests, in planta, showed a significant reduction of the frequency of both phenylpyrrole-resistant phenotypes, with a respective increase in the population of the wild-type strain of the pathogen.     

Involvement of FgMad2 and FgBub1 in regulating fungal development and carbendazim resistance in Fusarium graminearum

Resistance to carbendazim of Fusarium graminearum is conferred by point mutation in the β₂‐tubulin gene that plays an important role in spindle assembly. The spindle assembly checkpoint is a cellular surveillance system that is critical for maintaining genomic stability. Predicted protein Mad2‐ and Bub1‐encoding genes in F. graminearum (FgMad2 and FgBub1) were isolated and characterized. There was no difference in FgMad2 and FgBub1 expression levels between carbendazim‐sensitive and ‐resistant strains; however, after carbendazim treatment FgMad2 expression increased while FgBub1 expression stayed the same. Both the FgMad2 and FgBub1 deletion mutants became more sensitive to carbendazim. The FgMad2 deletion mutants grew more slowly, produced fewer conidia and both hyphae and conidia were malformed. Conversely, deletion of FgBub1 had no effect on fungal development other than a reduction in conidia production. FgMad2 deletion mutants exhibited a severe decrease in perithecia production and pathogenicity along with a down‐regulation of trichothecene production, whereas FgBub1 deletion mutants exhibited only a slight reduction in perithecia production and was accompanied by a twofold increase in trichothecene production. Overall, the results indicate that both FgMad2 and FgBub1 are involved in carbendazim resistance and trichothecene biosynthesis, and FgMad2 plays an important role in fungal development in F. graminearum.     

Vegetative compatibility grouping in Botrytis cinerea using sulphate non-utilizing mutants

Twenty-one strains of Botrytis cinerea isolated from six plant species on ten sites throughout Israel, as well as a strain from France, were tested for vegetative and mycelial incompatibility, pathogenicity, resistance to the fungicides carbendazim and iprodione, and colony morphology. Selenate-resistant mutants were isolated from the strains as spontaneous, fast-growing sectors arising from restricted colonies on medium amended with sodium selenate with a mean frequency of 0.04 sectors/colony; 81% of the sectors were sulphate non-utilizing (sul) mutants. One hundred and four sul mutants were divided into two complementary groups: resistant (66 mutants) and sensitive to chromate. Based on compatibility reactions between chromate-resistant and chromate-sensitive sul mutants, 12 strains were compatible only with themselves and were each classified as belonging to different vegetative compatibility groups (VCGs). Nine strains were each compatible with one to three other strains, and were assembled into three multi-member VCGs. Mycelial incompatibility between wild-type strains (barrage), in the form of a zone of dark pigmentation or sparse mycelium with or without dark pigmentation of the agar along the line of confrontation, was observed for 70% of the inter-strain pairings. There was no correspondence in compatibility between strains revealed by two approaches: strains in different VCGs did not necessarily produce a barrage. However, self-compatibility was observed both as heterokaryon formation between complementary sul mutants and as an absence of barrages between mycelia of wild-type strains; wild-type strains belonging to the same VCG did not exhibit strong barrages, although weak antagonistic reactions were observed. Strains in two multi-member VCGs showed the same patterns of resistance to carbendazim and iprodione; the third multi-member VCG contained isolates with different patterns of resistance. Four morphological types were revealed among wild-type strains: conidial (five strains), sclerotial (six strains), intermediate (ten strains), and mycelial (one strain). On bean leaves, conidial strains were more aggressive than sclerotial strains.     

Application of cycleave PCR to the detection of a point mutation (F167Y) in the β2‐tubulin gene of Fusarium graminearum

BACKGROUND: Resistance of Fusarium graminearum to the benzimidazole fungicide carbendazim is caused by point mutations in the β₂‐tubulin gene (FGSG_06611.3). The point mutation at codon 167 (TTT → TAT, F167Y) occurs in more than 90% of field isolates in China. It is important to find a suitable method for rapid detection and quantification of this point mutation in the F. graminearum populations. RESULTS: A pair of primers, Codon167F/Codon167R, were designed to amplify a fragment containing the mutation site, and two cycling probes labelled with different fluorescent reporters were used to detect whether the mutation was present. A cycleave real‐time PCR method was developed for rapid determination of the frequency of this point mutation in 282 F. graminearum perithecia collected from different fields in Jiangsu Province, China. The mutation frequency in ascospores from the perithecia to carbendazim by a spore germination assay was 6.0%, while the frequency of the point mutation at codon 167 by the cycleave real‐time PCR assay was 3.9%. CONCLUSION: The cycleave real‐time PCR method is suitable for accurate detection of the codon 167 point mutation. The frequency of this mutation in the β₂‐tubulin gene represents the resistance frequency in F. graminearum populations to carbendazim. Copyright © 2011 Society of Chemical Industry     

Detection of enzymatic activity and partial sequence of a chitinase gene in <Emphasis Type="Italic">Metschnikowia pulcherrima</Emphasis> strain MACH1 used as post-harvest biocontrol agent

Two antagonistic yeast strains Metschnikowia pulcherrima MACH1 and Rhodotorula sp. PW34 were tested for their efficacy against Botrytis cinerea in vitro and in vivo on apples. Metschnikowia pulcherrima strain MACH1 showed higher inhibition of B. cinerea compared to the strain PW34 in vitro on potato dextrose broth. Further, yeast strain MACH1 showed higher efficacy in reducing grey mould on apples compared to PW34 and the untreated control. In addition, partially purified extracellular proteins from strain MACH1 showed an inhibition to B. cinerea in vitro. The antagonistic yeast strains were tested for their efficacy to produce chitinases in different liquid media, including apple juice, amended with or without cell wall preparations (CWP) of B. cinerea. The study showed a higher production of chitinases from M. pulcherrima strain MACH1 when compared to PW34. Interestingly, the strain MACH1 secreted higher chitinases in the presence of cell wall fractions of B. cinerea. For this reason, the chitinase gene of strain MACH1 was amplified using PCR reactions and the nucleotide sequence data showed high homology to chitinases of other yeast strains. The results of the current study show that M. pulcherrima strain MACH1 has the ability to secrete chitinases in different liquid media including apple juice, and the enzyme could be involved in the post-harvest biological control of B. cinerea.     

Identification of a novel point mutation in the <Emphasis Type="Italic">β</Emphasis>-tubulin gene of <Emphasis Type="Italic">Botrytis cinerea</Emphasis> and detection of benzimidazole resistance by a diagnostic PCR-RFLP assay

The molecular basis of resistance to benzimidazole fungicides with laboratory and field mutant isolates of Botrytis cinerea was investigated. After chemical mutagenesis with N-methyl-N-nitrosogouanidine (NMNG) two different benzimidazole-resistant phenotypes were isolated on media containing carbendazim or a mixture of carbendazim and diethofencarb. The mutant isolates from the fungicide-mixture-containing medium were moderately resistant to carbendazim with wild-type tolerance to diethofencarb while mutant isolates from carbendazim-containing medium were highly resistant to carbendazim but sensitive to diethofencarb. The studied field isolates were highly resistant to benzimidazoles and sensitive to diethofencarb. Study of fitness characteristics of benzimidazole highly-resistant isolates showed that the resistance mutation(s) had no apparent effect on fitness-determining parameters. Contrary to this, the moderately benzimidazole-resistant strains, with no increased diethofencarb sensitivity, had a significant reduction in certain ecological fitness-determining characteristics. Analysis of the sequence of the β-tubulin gene revealed two amino acid replacements in the highly benzimidazole-resistant mutants compared to that of the wild-type parent strain. One was the glutamic acid (GAG) to alanine (GCG) change at position 198 (E198A), identified in both laboratory and field highly benzimidazole-resistant isolates, a mutation previously implicated in benzimidazole resistance. The second was a novel benzimidazole resistance mutation of glutamic acid (GAG) to glycine (GGG) substitution at the same position 198 (E198G), identified in a highly benzimidazole-resistant laboratory mutant strain. Molecular analysis of the moderately benzimidazole-resistant strains revealed no mutations at the β-tubulin gene. A novel diagnostic PCR-RFLP assay utilising a BsaI restriction site present in the benzimidazole-sensitive (E198) but absent in both resistant genotypes (E198G and E198A) was developed for the detection of both amino acid replacements at the β-tubulin gene.     

Genetic Analysis of the Role of Trichothecene and Fumonisin Mycotoxins in the Virulence of Fusarium

The phytotoxicity of the Fusarium trichothecene and fumonisin mycotoxins has led to speculation that both toxins are involved in plant pathogenesis. This subject has been addressed by examining virulence of trichothecene and fumonisin-nonproducing mutants of Fusarium in field tests. Mutants were generated by transformation-mediated disruption of genes encoding enzymes that catalyze early steps in the biosynthesis of each toxin. Two economically important species of Fusarium were selected for these studies: the trichothecene-producing species Fusarium graminearum, which causes wheat head blight and maize ear rot, and the fumonisin-producing species F. verticillioides, which causes maize ear rot. Trichothecene-non-producing mutants of F. graminearum caused less disease than the wild-type strain from which they were derived on both wheat and maize, although differences in virulence on maize were not observed under hot and dry environmental conditions. Genetic analyses of the mutants demonstrated that the reduced virulence on wheat was caused by the loss of trichothecene production rather than by a non-target mutation induced by the gene disruption procedure. Although the analyses of virulence of fumonisin-non-producing mutants of F. verticillioides are not complete, to date, the mutants have been as virulent on maize ears as their wild-type progenitor strains. The finding that trichothecene production contributes to the virulence of F. graminearum suggests that it may be possible to generate plants that are resistant to this fungus by increasing their resistance to trichothecenes. As a result, several researchers are trying to identify trichothecene resistance genes and transfer them to crop species.     

Characterisation of novel <Emphasis Type="Italic">Fusarium graminearum</Emphasis> microsatellite markers in different <Emphasis Type="Italic">Fusarium</Emphasis> species from various countries

Fifteen novel microsatellite markers were isolated from Fusarium graminearum. The level of polymorphism at these novel and 13 previously published microsatellite markers was analysed in 33 F. graminearum strains from Europe, North America, and Nepal. The number of alleles for each of the novel markers ranged from 4 to 20 and gene diversity from 0.417 to 0.962. In comparison with the previously published markers, the resolution for distinguishing among different strains was slightly increased. Twenty-seven markers were also detectable in three F. culmorum strains and one F. crookwellense strain. None of the markers was detected in three F. avenaceum and four F. poae strains, underlining the potential use of these microsatellite markers for species differentiation.