Benomyl and methyl-2-benzimidazolecarbamate (MBC): Biochemical,cytological and chemical aspects of toxicity to Ustilago maydis and Saccharomyces cerevesiae

Action of methyl-2-benzimidazolecarbamate (MBC), a breakdown product of benomyl, was studied in synchronous cultures of Ustilago maydis and Saccharomyces cerevesiae. Cells treated with MBC developed similarly to control cells until the two portions of the doublets (joined mother and daughter “cells”) were morphologically equivalent. At this point development of the treated cells ceased but control doublets separated to form two new cells. The compound did not prevent DNA synthesis during the first cell cycle of synchronized S. cerevesiae cells but did prevent completion of cell division, as it did also in synchronized U. maydis sporidia which already contained the DNA complement needed for completion of the first division. Mitosis did not go to completion in MBC-treated cells so the doublets formed contained only a single compact nucleus. The effect of MBC markedly resembles mitotic arrest caused by colchicine and isopropyl N-phenylcarbamate in higher plants and griseofulvin in the fungus Aspergillus nidulans. Inhibition of DNA synthesis in U. maydis and inhibition of cytokinesis in both organisms studied are secondary effects attributable to mitotic failure.A volatile compound toxic to U. maydis and S. cerevesiae was demonstrated in air over moistened benomyl. Hexane, through which such air samples were passed, or hexane extracts of aqueous suspensions of benomyl contained a substance having the same ir spectrum and retention time in two gas chromatographic systems as butyl isocyanate (BIC).BIC inhibited respiration of U. maydis and S. cerevesiae in a manner similar to benomyl. Benomyl partially inhibited respiration of subcellular particles from both organisms. MBC had essentially no effect on whole cell or subcellular particle respiration of either organism. Toxicity of benomyl preparations is not attributed to “benomyl” as such, but to two breakdown products, MBC and BIC. Therefore, differential effects of benomyl preparations and of MBC on growth and metabolism in fungi should be ascribed to BIC.     

Effect of benomyl on the potato cyst nematode,Heterodera rostochiensis

Benomyl is a strong fungistat against certain groups of fungi and it does not have much effect on most plant-parasitic nematodes.Heterodera spp., however, are affected, and the literature indicated a strong effect onHeterodera rostochiensis in the field. Laboratory tests demonstrated that hatching of larvae fromH. rostochiensis cysts is strongly inhibited by benomyl in vitro. In a pot trial, benomyl effectively suppressed formation of new cysts on the roots of potatoes grown in sand, clay soil or lightsandy loam. There was no such effect in soil rich in organic matter probably because of strong adsorption of benomyl. Control ofH. rostochiensis by benomyl in soil could be due to suppressed hatch or to other mechanisms, such as inhibition of larval penetration of roots. In view of its long persistence in the soil and of its various side-effects, benomyl does not seem very promising for practical control ofHeterodera rostochiensis.Samenvatting Benomyl is bekend als een middel met sterk fungistatische eigenschappen ten aanzien van bepaalde groepen schimmels. Het heeft in het algemeen weinig effect op planteparasitaire aaltjes. Cystevormende aaltjes echter, en in het bijzonderHeterodera rostochiensis, zijn, volgens aanwijzingen uit de literatuur gevoeliger voor benomyl.Onder laboratoriumomstandigheden bleek het uitkomen van de larven vanH. rostochiensis uit de cysten sterk door benomyl te worden geremd (Tabel 1 en 2). In een potproef werd door een grondbehandeling met benomyl de vorming van nieuwe cysten op aardappelwortels onderdrukt, afhankelik van de grondsoort (Tabel 3). Naarmate de grond meer organische stof of klei bevat, is het effect minder groot. Bijgebruik van rivierzand trad reeds bij een dosering van ongeveer 1 ppm benomyl een volledige on derdrukking van cystevorming op. Maar bij potgrond (59% organische stof) was er, ook bij een tien maal hogere dosering, geen effect. Verder onderzoek zal moeten uitwijzen of het effect in grond, berust op de remming van het uitkomen van larven uit de cysten, of (ook) op de beïnvloeding van andere aspecten van het parasitair gedrag van dit aaltje, zoals de penetratie in de wortels.Gezien de grote persistentie van benomyl in de grond, en de nevenwerkingen, lijkt het middel minder geschikt voor toepassing in de praktijk.Guest worker of TNO, Netherlands Organization of Applied Scientific Research.     

Inhibition of cutinase and prevention of fungal penetration into plants by benomyl—A possible protective mode of action

Cutinase purified from Fusarium solani f. sp. pisi was effectively inhibited by the fungicide benomyl. This irreversible inhibition was shown to be caused by the reaction of n-butylisocyanate, a breakdown product of benomyl, with the active serine in the enzyme. Methyl-2-benzimidazolecarbamate (MBC), the other breakdown product of benomyl, had no cutinase inhibitory activity. The fungicidal action of benomyl was measured by a bioassay using pea stems with or without intact cuticular and cell wall barriers. Benomyl prevented infection of Pisum sativum by conidia of F. solani f. sp. pisi only on intact pea stems with 50% efficiency at 7 μM, but was without effect on wounded stems or when benomyl (up to 50 μM) was applied 24 hr after inoculation with spores. Both MBC and benomyl strongly inhibited mycelial growth with 50% inhibition at 2 μM indicating a toxic mode of action. The prevention of fungal infection on only intact stem surfaces was established, however, with a fungal strain which was relatively tolerant to the toxic effect of these fungicides. With this strain complete protection was obtained at 5 μM benomyl which caused only 30% inhibition of growth. These results suggest that benomyl at low concentrations prevented fungal penetration and thus infection probably through inhibition of cutinase. It is also shown that treatment of cutin with n-butylisocyanate made it resistant to cutinase action.     

Effect of benomyl on soil fungi associated with rye. 2. Effect on fungi of culm bases and roots

The mycoflora of culm bases and roots of rye was assessed in field trials, where benomyl was applied at dose rates ranging from 0.24 to 4.80 kg ha^?1. Samples of culm bases were taken three times during the growing season, those of roots only at the harvest date. On culms with various symptoms from untreated plots,Pseudocercosporella herpotrichoides, Alternaria spp. andGerlachia nivalis were prevalent and on those from benomyl-treated plotsAlternaria spp. andFusarium culmorum. In later stages of growth,G. nivalis sharply declined andAlternaria spp. andF. culmorum increased. At the end of the season,Periconia macrospinosa andTyphula incarnata appeared in treated plots. In samples of roots taken more than two months after the last spray, porosporous dematiaceous species (Alternaria, Ulocladium andDendryphion), Mortierella spp. and other resistant fungi were prevalent in plots treated with 1.20 kg ha^?1 or more, but not in those that received 0.24 kg ha^?1, which is recommended for disease control. Lower counts were recorded only for species that are highly sensitive in vitro, e.g.Microdochium bolleyi andTrichoderma spp. Some fusaria were either not affected or tended to be slightly stimulated by the treatment. An attempt was made to attribute the incidence of these moderately sensitive fungi to the effect of the fungicide on non-pathogens.     

Phylogenetic relationship and fungicide sensitivity of members of the <Emphasis Type="Italic">Colletotrichum gloeosporioides</Emphasis> species complex from apple

The members of the Colletotrichum gloeosporioides species complex (CGSC), the dominant pathogens of apple bitter rot in Nagano prefecture, Japan, were reidentified and the relationship between the species and fungicide sensitivity was revealed. Based on phylogenetic analysis of the ApMat locus with the neighbor-joining (NJ) method, isolates from apple contained three species of the CGSC; C. fructicola, C. aenigma, C. siamense, and three clades of the CGSC: Clade V, S and K. Colletotrichum fructicola and Clade S dominated in Nagano Prefecture. Isolates of C. siamense, C. aenigma and Clade V, S and K remained sensitive to benomyl and quinone outside inhibitor (QoI) fungicides, while C. fructicola often developed resistance to benomyl and QoI fungicides. These results suggest that the development of fungicide resistance differs among members of the CGSC.     

Chemical control of Didymella bryoniae in cucumbers

The effect of a number of fungicides on mycelial growth ofDidymella bryoniae on agar was tested and compared with the effect on inoculated plants and in commercial crops. Activities in vitro and vivo are not always correlated. Spraying of inoculated young plants is a reliable and quick method for testing fungicides againstD. bryoniae.In commercial crops the treatments in decreasing order of effectiveness were weekly sprays of benomyl 0.025–0.05% active ingredient (a.i.), chlorothalonil 0.15% and triforine 0.02% a.i. These chemicals should be sprayed alternately to prevent the development of strains tolerant to benzimidazole derivates.Samenvatting Een aantal chemische middelen zijn in vitro en op verschillende manieren in vivo getoetst op hun werking tegenDidymella bryoniae. De resultaten in vitro zijn niet altijd positief gecorreleerd met die in vivo (Tabel 1 en 2). De resultaten na inoculatie van jonge plantjes komen wel overeen met die onder praktijkomstandigheden (Tabel 3, 4 en 5). Op jonge plantjes kunnen dus middelen op een snelle wijze en in een beperkte ruimte betrouwbaar worden getoetst.Onder praktijkomstandigheden werd het beste resultaat verkregen met benomyl 0,025–0,05% actieve stof, zowel wat betreft de gewas- als de vruchtaantasting. Hierop volgde chloorthalonil 0,15% a.s. en triforine 0,02% a.s. (Tabel 4 en 5). IndienD. bryoniae met chemische middelen moet worden bestreden, is het noodzakelijk dat er bijna elke week wordt gespoten (Tabel 5). Teneinde het ontstaan van resistentie van schimmels tegen benomyl en andere MBC-producerende fungiciden te voorkomen, wordt aanbevolen om benomyl, chloorthalonil en triforine afwisselend toe te passen. Met dit schema worden vele andere schimmelziekten in komkommer eveneens bestreden.     

Effect of waterborne benomyl on the hematological and antioxidant parameters of the Nile tilapia, Oreochromis niloticus

The present study was conducted to determine the 96 h-LC₅₀ of benomyl to the Nile tilapia, Oreochromis niloticus and to investigate the biochemical or hematological indices of blood and the alterations in the antioxidant enzymes of this fish in response to sublethal concentrations of benomyl. Fish weighing 71.61 ± 12.05 g were used in this study; they were subjected to fasting for 4 weeks before treatment. An aqueous solution of benomyl (0, 0.5, 1, 2, 4, 8, and 16 mg L⁻¹) was administered for 96 h to determine the LC₅₀. The 96 h-LC₅₀ value of benomyl was 4.39 (3.23-5.60) mg L⁻¹ in the present study. For 5 weeks, the aqueous solution of benomyl (0, 100, 200, and 400 μg L⁻¹) was administered to investigate its effect on the hematological parameters and antioxidant enzymes. The predominant hematological findings in fish exposed to benomyl were as follows: no significant change in the Hb (g dL⁻¹) level, MCV (μm³), MCH (pg) and MCHC (%) as compared to the control. Benomyl exposure led to greater increases in the GPT, GOT (Karmen-unit), LDH (Wroblewski unit), total cholesterol, Fe, and Ca (mg dL⁻¹) values, whereas the levels of ALP (KA unit), total protein, triglyceride, albumin, and Mg (mg dL⁻¹) did not increase. Benomyl increased the in vivo HSI (%), GST (nmol min⁻¹ mg protein⁻¹), and SOD (U mg protein⁻¹) values in the fish livers in the test group, unlike those in the control group for 5 weeks. At concentrations higher than 100 μg L⁻¹, benomyl affected the GST and SOD levels of Nile tilapia in a dose- and time-dependent manner. The present findings suggest that the in vivo hepatotoxicity associated with benomyl may, in part, result from the hematological index, and antioxidants may provide limited protection against benomyl toxicity.     

Interference of benomyl and methyl-2-benzimidazolecarbamate (MBC) with DNA metabolism and nuclear divisions in Fusarium oxysporum

Methyl-1-(butylcarbamoyl)-2-benzimidazolecarbamate (benomyl) severely decreased DNA synthesis when applied at 3.5 × 10^?6M during the G1 phase of germinating conidia of Fusarium oxysporum; nuclear divisions were completely inhibited at a fungicide concentration of 10 × 10^?6M. The same concentration applied only after the S phase also completely inhibited the nuclear divisions. This dual interference of benomyl with DNA formation and mitosis might be related to a disturbed phosphorus metabolism.     

Techniques for isolatinghelminthosporium pedicellatum from infected corn roots

In a comparison of potato dextrose agar (PDA) and Czapek’s Dox broth (CDB) agar with and without benomyl, aa growth media forHelminthosporium pedicellatum Henry (Trichometasphaeria pedicellatum Nelson), PDA was 1.5 times more efficient than CDB under all conditions. Growth rate was negatively correlated with benomyl concentration, and was inhibited 11% and 74% at benomyl concentrations of 10 μg and 200 μar, respectively, on both media. Pure isolates of the fungus were recovered from corn roots only where roots had been sterilized with 10% sodium hypochlorite. Root sterilization may thus be more important than medium amendment in isolating pure cultures ofedicellatum from plant roots.     

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.     

A comparison of the in vitro antifungal spectra of thiophanates and benomyl

The antifungal spectra of thiophanate, thiophanate-methyl and its derivative 2-(3-methoxycarbonyl-2-thioureido) aniline (NF 48), were similar to that of benomyl. The order of effectivenessin vitro was: benomyl>NF 48> thiophanate-methyl > thiophanate, benomyl being by far the most and thiophanate by far the least active compound.The effect on mycelial growth of two fungi with an irregular inhibition pattern are presented in detail, viz. ofColletotrichum acutatum andGliocladium roseum. In the first case mycelial growth was inhibited to over 50% at low concentrations; the inhibition, however, was not further enhanced with increasing concentrations. Mycelial growth ofGliocladium roseum was maximally suppressed at low concentrations of the fungicides, whereas in this case an increase of the concentrations resulted in a decrease of inhibitory activity until a constant level had been reached. Conidiobolus eurymites, unlike the other Zygomycetes tested, proved to be sensitive to the four fungicides.Samenvatting Het fungitoxisch spectrum van de systemische fungiciden thiofanaat, thiofanaat-methyl en een derivaat hiervan, NF 48, bleek gelijk te zijn aan dat van benomyl.In vitro was de volgorde in remmende werking op de myceliumgroei: benomyl > NF 48> thiofanaat-methyl > thiofanaat. Van deze verbindingen was benomyl verreweg het meest en thiofanaat duidelijk het minst fungitoxisch. Thiofanaatmethyl en NF 48 gaven in dit opzicht slechts weinig verschil te zien.De invloed van de fungiciden op de myceliumgroei van twee schimmels met een onregelmatig remmingspatroon is weergegeven in de Fig. 1 en 2. Bij geringe concentraties van de fungiciden in het medium werd de myceliumgroei vanColletotrichum acutatum reeds tot ruim 50% geremd. Verdere toename van de fungiciden, zelfs tot zeer hoge concentraties (1000 M), had echter geen sterkere remming ten gevolge. BijGliocladium roseum bleek de myceliumgroei het sterkst geremd bij lage concentraties (bv. voor benomyl bij ca 4 M). Verhoging van de concentraties had een afname in remmende werking ten gevolge tot een niveau van 75% remming was bereikt.In tegenstelling tot de andere Zygomyceten, die tot nu toe op gevoeligheid ten aanzien van benomyl onderzocht werden, bleekConidiobolus eurymites, een saprofyt uit de bodem, gevoelig te zijn voor alle vier fungiciden.     

Benomyl-resistantCercosporidium personatum in peanut fields in the Sharon region of Israel

Cercosporidium personatum (Berk. & Curt.) Deighton was sampled from five peanut fields in the Sharon region of Israel, where benomyl failed to control late leafspot. Most samples were resistant to 10 ppm carbendazimin vitro.     

A selective medium for the detection ofAlternaria dauci andAlternaria radicina

A selective medium to detectAlternaria dauci andA. radicina on seed, plant debris and other substrates was developed. Growth and sporulation by most undesired organisms was reduced, but adequate mycelial growth and sporulation byA. dauci andA. radicina was maintained so they could be identified by their unusual mycelial growth or characteristic spores. The medium is based upon carrot leaf extract, which promotes profuse sporulation by both pathogens. Glucose, sodium polypectate and mineral salts further enhanced spore and mycelial production. Streptomycin sulfate and metalaxyl (or mefenoxam), combined with either benomyl or thiophanate-methyl, reduced growth and sporulation of unwanted organisms. There were strong interactions among the fungicides, bactericide, and most other medium components. There were also significant effects on the type of mycelial growth produced by some medium components. If the unique, dark mycelia produced by both pathogens is the desired detection method, the medium components must be optimized for eitherA. dauci orA. radicina because the two fungi responded differently in this regard. All medium configurations allowed both fungi to be identified by their characteristic spores. The sensitivity of theAlternaria dauci-radicina selective medium (ADRSM) to detectA. radicina on carrot seeds was similar to other methods presently in use, but ADRSM was more sensitive than the other methods for detectingA. dauci on infested carrot seeds.     

The effect of fungicides on fungal antagonists of onion white rot and selection of dicarboximide-resistant biotypes

The sensitivity of four fungal antagonists (Chaetomium globosum, Trichoderma harzianum, T. viride, Trichoderma sp.) to six fungicides was evaluated. The fungi were insensitive to captan, mancozeb and thiram but were sensitive to benomyl (EC₅₀ < 0 3/μg/ml) and the two dicarboximides iprodione and procymidone (EC₅₀ < 3 3/μg/ml). Conidia and mycelia of the four fungi were exposed to a series of ultraviolet irradiations in order to select biotypes resistant to benomyl and iprodione. Resistance to benomyl was not induced but a number of biotypes of each fungus were isolated from irradiated cultures that showed resistance to iprodione (EC₅₀ values of up to 56·4, 177·8, 171·5 and 216·4/μg/ml for Chaetomium globosum, Trichoderma harzianum, T. viride and Trichoderma sp., respectively). With three exceptions, all selected biotypes retained their fungicide-resistance after being extensively cultured on fungicide-free medium. In general, growth of the fungicide resistant biotypes on MEA plates and in MYE liquid medium was equal to that of the corresponding wild-type strains. One resistant biotype of Trichoderma sp. (C62UV3) grew significantly (P < 0·05) better both on solid and in liquid medium. This resistant biotype was also observed to have greatly enhanced conidial production on agar plates. All fungicide resistant-biotypes retained the ability to antagonize the pathogen Sclerotium cepivorum in dual culture and, in two instances (A53UV1, A53UV5), exhibited greater antagonism as evidenced by the production of larger inhibition zones. Similarly, with two exceptions, the resistant biotypes retained the ability to control onion white rot disease caused by S. cepivorum in the glasshouse. In particular, two fungicide-resistant biotypes (Trichoderma sp. C62UV3 and Chaetomium globosum A53UV1) reduced white rot of onion more effectively than did their respective wild-type parents.     

Evaluation of systemic fungicides for use in the glasshouse industry

The primary evaluation of ten fungicides for the control of cucumber powdery mildew (Sphaerotheca fuliginea), tomato leaf mould (Cladosporium fulvum) and tomato grey mould (Botrytis cinerea) indicated that nine of them were systemically active against one or more of the pathogens when applied as soil drenches. Benomyl, thiophanate, thiophanate-methyl, thiabendazole and Cela W52A showed activity against all three diseases, but dimethirimol, triarimol and dodemorph were more specific and mainly active against cucumber powdery mildew. An in vitro bioassay generally indicated that fungitoxicity was not translocated from sprayed to unsprayed leaves, and though toxicity to B. cinerea, following soil drenches of benomyl, thiophanate and thiophanate-methyl, was detected by bioassay in leaf sap and calyces it was not detected in fruit. Furthermore, extended evaluation of benomyl for the control of grey mould on cropping tomotoes showed that in the presence of abundant B. cinerea inoculum, benomyl soil drench treatment effectively controlled flower and leaf infection but not fruit ghost-spotting.     

Ultrastructural and histoautoradiographic study of benomyl action on microconidia of Fusarium oxysporum in germination

Ultrastructural changes produced by benomyl, a benzimidazole derivative, during the first 48 hr of germination of Fusarium oxysporum f. sp. melonis conidia are investigated; an attempt to localize ³H-benomyl by autoradiography was carried out. Benomyl has fungicidal and fungistatic activity. Indeed it causes the lysis of 10% of the germinating microconidia and reduces drastically the growth of the germ tube in the other cells. In this latter case, cell wall thickening and anomalous morphology have been observed. Lysosome-like vesicles appear in the cytoplasm. It is proposed that benomyl acts on membrane integrity.     

稻田土壤8种真菌对4种杀菌剂的敏感性分析

基于ITS-5.8s rDNA区序列,初步鉴定了浙江大学试验基地稻田土壤的8种常见真菌。采用菌丝生长速率法测定了井冈霉素、多菌灵、咪鲜胺和三唑酮4种常用杀菌剂对这8种土壤真菌的抑菌活性,以及多菌灵与咪鲜胺混用对禾柄锈菌、黑曲霉和长枝木霉3种真菌菌丝生长的抑制作用。结果表明:8种真菌对咪鲜胺最敏感,EC50值在0.091~3.1 mg/L之间;多菌灵的EC50值在0.15~1.8 mg/L之间;三唑酮的EC50值在0.57~8.5 mg/L之间;井冈霉素的敏感性最低,EC50在12~94 mg/L之间。由4种杀菌剂的SSDs曲线分析结果得出,不同真菌对杀菌剂敏感性存在显著差异。当多菌灵与咪鲜胺按其EC50值分别以体积比3∶2、1∶1和7∶3混用时,增效系数分别为1.615、1.039和1.042,表现出一定的协同或相加作用。本研究结果为进一步阐明杀菌剂对稻田生态系统真菌群落的生态效应及杀菌剂的科学合理使用提供了重要依据。     

Action of benzimidazole fungicides on resorcinolic lipid metabolism in rye seedlings depends on thermal and light growth conditions

The content and homologue composition of alkylresorcinols were investigated in rye seedlings (Secale cereale L.) treated with benzimidazole fungicides and grown for 5 days under various thermal and light conditions. The fresh and dry biomasses of green and etiolated plants were greatly increased by benomyl and carbendazim at 29 °C. At 22 and 15 °C benomyl had an inhibitory action on growth of rye, whereas the effects caused in the presence of carbendazim were slightly dependent on external stimuli. On the other hand, benomyl enhanced resorcinol biosynthesis in green seedlings grown at all temperatures, while carbendazim only at 29 °C. In plants kept in the darkness, both fungicides increased content of alkylresorcinols at 29 °C and decreased its at 22 and 15 °C. The qualitative pattern of resorcinolic homologues was also significantly modified in the presence of benomyl and carbendazim and depended on other physical stimuli.     

Systemic fungicides in strawberry: field control of fruit rot (Botrytis cinerea pers.) and residue study

In a detailed study of the effect of weekly sprays with benzimidazole-derivative fungicides on strawberry plants, cv. Tioga, higher yields were recorded, as well as reduced fruit rot caused by the fungusBotrytis cinerea Pers. The two fungicides tested, benomyl and flowable and soluble formulations of TBZ, gave similar control of fruit rot. The residue level found in leaves and fruits was about the same throughout the yielding season. Residue levels of 300-433 ppm TBZ (flowable form) and 94-110 ppm benomyl were detected in the leaves, whereas the levels in the fruit were 6.4-8.2 ppm and 1.5-3.8 ppm for the two compounds, respectively. Two weeks after the last spray application, the residue level of TBZ in teaves was reduced by 50%, and of benomyl by only 20%.     

Further studies of the inheritance of benomyl resistance in Venturia pirina isolated from pear orchards in Israel

Fourteen single-spore cultures of benomyl-resistant Venturia pirina were isolated from pear scab lesions at four sites in Israel. According to the ability of the isolates to germinate and grow at varying benomyl concentrations, four levels of resistance were determined in vitro : three isolates with low resistance (LR) grew at 0.5 but not at 5 μg/ml benomyl: five moderately resistant (MR) isolates grew at 5 but not at 50 μg/ml benomyl: five highly resistant (HR) isolates grew at 50 μg/ml but their hyphae were curled: and one isolate with very high resistance (VHR) grew unaffected at 50 μg/ml benomyl. The difference between the HR and the VHR phenotypes was clearly shown on medium amended with N -(3.5-dichlorophenyl) carbamate (MDPC): only the VHR isolate showed negative cross-resistance to 1 μ g/ml MDPC. whereas HR isolates grew unaffected. Crosses between resistant isolates and sensitive wild types, as well as between different resistant isolates, showed that the various levels of resistance are conferred by four allelic mutations that constitute a polymorphic series at a single locus.