Biochemical and molecular mechanisms of diafenthiuron resistance in the whitefly,Bemisia tabaci

B-biotype Bemisia tabaci has developed high levels of resistance to many insecticides. To investigate the risks and explore possible mechanisms of resistance to diafenthiuron in B. tabaci, a 32.8-fold diafenthiuron-resistant strain (R-DfWf) was established after selection for 36 generations compared with the susceptible strain (S-Lab). Biochemical assays showed that the activity of cytochrome P450 towards p-NA was significantly higher (4.37-fold higher) in the R-DfWf strain than in the S-Lab strain. Similarly, the carboxylesterase (COE) activity and glutathione S-transferase (GST) activity were also significantly higher (3.12- and 1.83-fold higher, respectively) in the R-DfWf strain than in the S-Lab strain. The expression of five of seven P450 genes was significantly higher (>3-fold) in the R-DfWf strain than in the S-Lab strain. The expression of COE2 was significantly higher (>2.5-fold) in the R-DfWf than in the S-Lab strain. The expression of GST and GST2 was significantly higher (>2.3-fold) in the R-DfWf than in the S-Lab. Thus, cytochrome P450, COE and GST may appear to be responsible for the resistance to diafenthiuron in B. tabaci. It is also valuable for usage of insecticides for resistance management and control of this species.     

A single gene transfer of gibberellin biosynthesis gene cluster increases gibberellin production in a Fusarium fujikuroi strain with gibberellin low producibility

Fusarium fujikuroi, the causative agent of bakanae disease in rice, produces many kinds of secondary metabolites. Recently, two phylogenetic subgroups (F and G groups) of Japanese F. fujikuroi have been identified and found to have differences in their gibberellin (GA) and fumonisin production. G-group F. fujikuroi produces large amounts of GA, but is a fumonisin nonproducer. F-group produces large amounts of fumonisin, but is a GA low or nonproducer. We investigated the cause of low GA production in the F-group. Genetic mapping suggests that low GA production in the F-group strain Gfc0825009 is due to a GA gene cluster for GA biosynthesis. Analysis of the nucleotide and amino acid sequences of the genes in the GA gene cluster showed >98.4% homology between the F-group strain Gfc0825009 and the G-group strain Gfc0801001. Following a 7-day culture under low nitrogen conditions, we found that expression of P450-1, P450-4, and P450-2 in the cluster increased in the G-group strain and not in the F-group strain. We hypothesized that complementation by GA genes in the G-group strain would be required to increase GA production in the F-group strain. However, we found that this occurred with a single gene complementation of DES, P450-1, P450-4, or P450-2. Simultaneous increase in the expression of P450-1, P450-4, and P450-2 were detected in the complementary transformants. Moreover, the same phenomenon was observed by reintegration of its own P450-1. Our results suggest the presence of unknown regulatory mechanisms of the GA gene cluster in F. fujikuroi.     

Effect of fungal metabolites and some derivatives against Tribolium castaneum (Herbst) and Nezara viridula (L.)

Two fungal metabolites, aspyrone (3-(1,2-epoxypropyl)-5-hydroxy-6-methyl-5,6-dihydropyran-2-one) and asperlactone (3-(1,2-epoxypropyl)-5-(1-hydroxyethyl)-5-furan-2-one) were isolated from an Aspergillus ochraceus Wilhelm strain showing IGR activity against Tribolium castaneum (Herbst). Synthetic derivatives of aspyrone were produced using published methods. These derivatives together with aspyrone and asperlactone were tested for insect growth-regulating activity against T. castaneum, and for ovicidal activity against Nezara viridula L. Of the compounds tested asperlactone appeared to be the most active.     

Biological activity of neem seed kernel extracts and synthetic azadirachtin against larvae of Plutella xylostella L

The activity of two neem extracts, AZT and NEEM-AZAL (containing 30 and 3 mg azadirachtin ml^?1 respectively) and synthetic azadirachtin (AZ) against second-instar larvae (L2) of Plutella xylostella L. was examined using leafdip bioassays. On Chinese cabbage, AZ was significantly (P <0.05) less toxic (3 to 4-fold; LC₅₀ 0.54 μg AZ ml^?1) than either neem extract against a laboratory strain of P. xylostella (FS). The LC₅₀ values for AZT against the FS and another laboratory strain (Wellcome) were not significantly different on Chinese cabbage. The activity of AZT against the FS and Wellcome strains was similar on Chinese cabbage and Brussels sprout. AZT was significantly less toxic (3-fold) on Brussels sprout against an acylurea-resistant field strain (Sawi) when compared with the FS strain on Chinese cabbage. Larval mortality (at day 13) was found to increase with increasing exposure time of P. xylostella (FS) larvae to AZT-treated Chinese cabbage, although there was little difference in mortality between 48 and 120 h exposure. When AZT, NEEM-AZAL and AZ were applied at a dose (1 μg AZ ml^?1) which gave end-point mortalities between 50 and 90% (at day 13), all treatments delayed the development of a proportion of surviving larvae but no morphogenetic abnormalities were observed in larvae which reached pupation. Evidence for antifeedant (reduced weight gain) and repellant effects (choicechamber) for AZT were observed with L2 P. xylostella (Wellcome) on Chinese cabbage. AZT was also shown to have ovicidal activity against P. xylostella (Wellcome) at relatively high dose ranges (10-1000 μg AZ ml^?1) as well as some contact activity (FS strain) in topical bioassays. In residual bioassays on glass with adults of the hymenopteran endo-larval parasitoid of P. xylostella, Diadegma semiclausum (Ichneumonidae), AZT showed little or no activity at rates up to 1000 μg AZ ml^?1. In medium-volume (MV, 200 litre ha^?1) and ultra-low-volume (c. 1 litre ha^?1) spray bioassays on Brussels sprout, AZT gave 16-92% and 88-100% mortality respectively (Wellcome strain) at rates approximating to 1-20 g AZ ha^?1. The residual activity of AZT and NEEM-AZAL against P. xylostella (FS) on Brussels sprout (MV spray) was observed to decrease appreciably after three days, the decline in activity being particularly marked for NEEM-AZAL.     

Toxicology and metabolism of chloromethiuron in Boophilus microplus larvae

The toxicity of the acaricide chloromethiuron, 3-(4-chloro-o-tolyl)-1,1-dimethyl- (thiourea), and of nine related compounds to Boophilus microplus larvae was determined by a spray-tower method. Four of these compounds were toxic but only chloromethiuron and its N-demethyl derivative were of practical importance. Metabolism of [¹⁴C]chloromethiuron, in the formamidine-susceptible but organo-phosphorus-resistant Mt. Alford strain, was compared with that in a chlordimeform-selected Mt. Alford strain, which in laboratory tests was two to three times resistant to chloromethiuron, chlordimeform and amitraz. The latter strain produced smaller quantities of the toxic N-demethyl derivative than the Mt. Alford strain; this was the only resistance mechanism determined. Rates of degradation of chloromethiuron were the same in both strains. Piperonyl butoxide strongly antagonised the toxicity of chloromethiuron by 18 to 33 times and depressed the production of the N-demethyl derivative in both strains (0.3 times that of the control), while degradation rates of chloromethiuron itself were halved by piperonyl butoxide in both strains. These results indicated that the parent material was not toxic until oxidised to the N-demethyl derivative. As, in addition, some symptoms of chloromethiuron toxicosis in larvae were similar to those caused by formamidine acaricides, a common mode of lethal action is suggested.     

Inhibition of crown gall formation by Agrobacterium radiobacter biovar 3 strains isolated from grapevine

Graft unions of nursery stock of grapevine (Vitis vinifera L.) collected in Japan yielded pathogenic and nonpathogenic strains of Agrobacterium. On the basis of classical diagnostic tests, a sequence analysis, and a multiplex polymerase chain reaction method previously reported, the pathogenic strain was identified as Agrobacterium tumefaciens biovar 3, whereas the nonpathogenic strains were assigned to Agrobacterium radiobacter biovar 3. Stems of tomato (Lycopersicon esculentum Mill.) seedlings were inoculated with both A. tumefaciens biovar 3 strain G-Ag-27 as a pathogen and one of the control strains isolated from grapevine or A. radiobacter biovar 2 strain K84 as competitors to assay the suppression of gall formation caused by the pathogen. In a test with a 1 : 1 pathogen/nonpathogen cell ratio, all A. radiobacter biovar 3 strains reduced gall incidence and size compared to that of the positive control inoculated only with the pathogen. Strain VAR03-1 was especially effective in reducing the incidence of gall formation on grapevine and reduced gall size by 84%–100% of those on the positive control. Many tested nonpathogenic biovar 3 strains were bacteriocinogenic, causing an inhibition zone against A. tumefaciens biovar 3 strains on YMA medium. Strain VAR03-1 was the most effective against indicator strains and appears to be a promising agent for controlling crown gall of grapevine.     

GA1-2菌株的分离鉴定及其对香蕉尖孢镰刀菌的抑菌效果

为了从土壤中分离筛选对香蕉尖孢镰刀菌具有良好拮抗作用的放线菌,采用平板稀释涂布法从四川省会理县干热河谷小麦根际土壤中进行放线菌分离,并采用平板对峙法和孢子萌发法进行筛选,通过形态特征、培养特征、生理生化特征以及16S r DNA序列分析对筛选菌株进行鉴定。结果表明,从四川省会理县干热河谷小麦根际土壤筛选获得一株对香蕉尖孢镰刀菌4号生理小种Fusarium oxysporum f.sp.cubense race 4(FOC4)菌丝和孢子萌发都有很强抑制作用的菌株GA1-2,对FOC4菌丝和孢子萌发抑制率分别为36.34%和94.81%。菌株GA1-2与薰衣草灰链轮丝菌Streptoverticillium lavenduligriseum的亲缘关系最近,相似率达99.85%,且其形态特征、培养特征、生理生化特征也与薰衣草灰链轮丝菌基本相符,因此将菌株GA1-2初步鉴定为薰衣草灰链轮丝菌。     

NPTⅡ基因和GUS基因在小麦遗传转化中的应用

对小麦遗传转化上常用的选择标记基因NPTⅡ的选择剂及GUS基因在幼胚愈伤组织中的转移进行了研究。结果表明用G418作为选择剂,浓度为20-30mg／L较为合适。幼胚愈伤组织与农杆菌共培养3d后,即可检测到GUS基因的瞬时表达。农杆菌LBA4404由500 mg／L羧卞青霉素抑制;EHA105、AGLI由500 mg／L头孢霉素抑制。而不能用卡那霉素。     

黄芪根腐病生防放线菌筛选鉴定及其优化培养

为获得对黄芪根腐病具有强抗菌活性的生防菌株,采用平板对峙法、抑制菌丝生长速率法对分离于河西走廊盐碱土中的172株放线菌进行拮抗菌的筛选,获得优菌株DA4-4-1,根据其形态、培养特征、生理生化特性和16S rDNA基因序列进行系统发育学分析鉴定,并根据单因素试验与正交试验对培养基条件进行优化。结果表明,菌株DA4-4-1对黄芪根腐病的2种主要致病菌尖孢镰刀菌Fusarium oxysporum和茄腐镰刀菌F.solani均有较好的抑制作用,抑菌圈直径最大分别为20.16 mm和19.33 mm,抑菌率最大分别为34.62%和32.44%。结合16S rDNA基因序列的系统发育分析,将菌株DA4-4-1鉴定为链霉菌属蓝紫链霉菌Streptomyces tuirus。培养基最佳碳源为麦芽糖,氮源为(NH_4)_2SO_4,其优化条件为麦芽糖15 g/L、(NH_4)_2SO_4 0.5 g/L、NaCl 10 g/L、pH 7,在此条件下菌株DA4-4-1的抑菌率可达到86.72%。表明菌株DA4-4-1是1株具有较强抗菌活性的放线菌,具有良好的生物防治和开发应用潜力。     

Comparative toxicity of two ecdysteroid agonists,RH-2485 and RH-5992, on susceptible and pyrethroid-resistant strains of the Egyptian cotton leafworm,Spodoptera littoralis

The comparative toxicity of two non-steroidal ecdysteroid agonists, RH-2485 and RH-5992 (tebufenozide), on development stages, fecundity and egg viability of a susceptible laboratory strain and a pyrethroid-resistant field strain ofSpodoptera littoralis (Boisduval) was evaluated. Taking the LC₅₀s as the criterion, RH-2485 was 3–7-fold more potent than RH-5992 against the susceptible and 7–14-fold more against the field strain. The LC₅₀ of RH-2485 in the 1st and 6th instars of the susceptible strain was 0.32 and 0.57 mg a.i./l, respectively. The field strain showed a mild cross-resistance of about threefold to both compounds in 1st instars and to a lesser extent in 6th instars. A considerable increase in fecundity (~3-fold) and no effect on egg viability was observed when 6th instars were fed on cotton leaves treated with 0.25 mg a.i./l RH-2485 (~LC₄₀). Our results indicate that both compounds are potentially potent insecticides for controllingS. littoralis larvae, being 10-60-fold more potent than a previous ecdysteroid agonist, RH-5849.     

Multiple Resistance in the Larger House Fly Musca domestica in Germany

Populations of the housefly Musca domestica isolated from farms in different German districts with strong resistance problems were compared to laboratory strains with varying resistance spectra. Resistance against pyrethroids, organophosphates and carbamates was tested using impregnated filter papers, and by topical application using a susceptible housefly strain (origin WHO) for comparison. The multi-resistant fly strains tested had a strong resistance against these insecticide groups, ranging from 37- to >10000-fold for organophosphates and 150- to >6600-fold for pyrethroids. The constituent enantiomer pairs of the α-cyano-pyrethroid cyfluthrin were tested, as was beta-cyfluthrin. With respect to multi-resistant fly strains, the isomers II and IV had the best activity, with LD₅₀ values of 0·012 and 0·014 μg per fly, respectively. In addition, different groups of insect growth regulators (juvenile hormone analogues, chitin synthesis inhibitors and one triazine derivative) were tested in a special larvicidal test. The chitin synthesis inhibitors were quite effective against multi-resistant M. domestica strains except for one strain with strong resistance against chitin synthesis inhibitors, developed after extensive treatments with benzoylphenylureas for several years. The fly strains tested were not resistant against cyromazine. Additionally, the insecticides were combined with the synergists piperonyl butoxide, tributylphosphorotrithioate (DEF) and Cibacron blue and tested against the fly strain with the strongest resistance spectrum (‘Grimm’) in comparison to the susceptible strain (‘WHO-N’). Piperonyl butoxide had the greatest effect on the efficacy of cyfluthrin followed by Cibacron blue and DEF. In a parallel investigation with susceptible and resistant house fly strains, different enzyme activities related with resistance mechanisms were tested, e.g. glutathione S-transferase (3·5-fold) and mixed-function oxidase (2·3-fold). Implications of these results for management of insecticide resistance in M. domestica are discussed.     

Biological control of grapevine crown gall by nonpathogenic <Emphasis Type="Italic">Agrobacterium vitis</Emphasis> strain VAR03-1

A nonpathogenic strain of Agrobacterium vitis VAR03-1 was tested as a biological control agent against crown gall of grapevine (Vitis vinifera L.). A mixture of the nonpathogenic strain VAR03-1 and a tumorigenic strain G-Ag-27 of A. vitis at cell ratios of 1 : 1, 3 : 1, 9 : 1, and 99 : 1 significantly inhibited gall formation and size on stems of tomato (Lycopersicon esculentum Mill.). Strain VAR03-1 also inhibited gall formation on stems of both tomato and grapevine at a 1 : 1 cell ratio with several tumorigenic A. vitis strains isolated from different fields of grapevine in Japan. In biological control tests, when roots of grapevine and tomato seedlings were soaked in a cell suspension of strain VAR03-1 for 24 h before a 1-h soaking in a cell suspension of the pathogen and subsequent planting in pots of infested soil, strain VAR03-1 significantly reduced the incidence of gall formation on both plants.     

Effects of radiation on the fitness,sterility and arbovirus susceptibility of a Wolbachia-free Aedes albopictus strain for use in the sterile insect technique

BACKGROUND

The sterile insect technique (SIT) is a green and species-specific insect pest control technique that suppresses target populations by releasing factory-reared, radiosterilized males into the wild. Once released, it is important to be able to distinguish the released males from the wild males for monitoring purposes. Several methods to mark the sterile males exist. However, most have limitations due to monetary, process efficiency, or insect quality. Aedes albopictus is naturally infected with Wolbachia at a high prevalence, therefore the elimination of Wolbachia can serve as a biomarker to distinguish factory-reared male mosquitoes from wild conspecifics.

RESULTS

In this study, a Wolbachia-free Ae. albopictus GT strain was developed and its fitness evaluated, which was found to be comparable to the wild GUA strain. In addition, GT male mosquitoes were irradiated at the adult stage and a dose of 20 Gy or more induced over 99% sterility. Moreover, a dose of 30 Gy (almost completely sterilizing male and female mosquitoes) had limited effects on the mating competitiveness of GT males and the vector competence of GT females, respectively. However, radiation reduced mosquito longevity, regardless of sex.

CONCLUSION

Our results indicate that the Ae. albopictus GT strain can be distinguished from wild mosquitoes based on Wolbachia status and shows similar fitness, radio-sensitivity and arbovirus susceptibility to the GUA strain, indicating that it is feasible to use the GT strain to suppress Ae. albopictus populations for SIT programmes. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

Aerial Contamination of Sugarcane in Guadeloupe by Two Strains of Xanthomonas albilineans

Two sugarcane plots were set up in Guadeloupe with disease-free tissue cultured plants in a banana growing location distant from sugarcane fields. Thirteen weeks after planting sugarcane in the field, a Xanthomonas albilineans strain belonging to serotype 3 (strain XaS3) was detected in water sampled at sunrise on the leaves in the first plot. This strain randomly invaded the sugarcane canopy. Seven weeks later, a new strain belonging to serotype 1 (strain XaS1) appeared on leaves and populations of strain XaS1 progressively increased on the leaf surface, whereas populations of strain XaS3 progressively decreased. Leaf scald symptoms were first noted 26 weeks after sugarcane planting. However, only strain XaS1 was isolated from leaves and a few sugarcane stalks showing symptoms. Both strains also colonized the second field plot, which was studied at the end of the experiment to avoid human interference of aerial contamination of sugarcane. After inoculation of three sugarcane cultivars by the decapitation technique, strain XaS1 was as virulent or more virulent than five other strains of X. albilineans isolated from diseased sugarcane plants in Guadeloupe. Although strain XaS3 colonized a few stalks, it failed to produce any symptoms and was the least virulent strain. Leaf surface colonization by X. albilineans was reproduced in a greenhouse trial by spraying the pathogen on sugarcane foliage. After 8 weeks, the pathogen was isolated from disinfected leaf blades. Although the leaf scald pathogen is thought to be mainly transmitted by infected cuttings, aerial transmission of X. albilineans is also known to occur. These results indicate the importance of sugarcane phyllosphere colonization by virulent strains in the epidemiological cycle of leaf scald disease in Guadeloupe.     

Laboratory selected resistance to diflubenzuron in larvae of aedes aegypti

The variation in tolerance to diflubenzuron [1-(4-chlorophenyl)-3-(2,6-difluorobenzoyl)urea] was examined in fourth instar larvae of seven strains of Aedes aegypti, some of which were resistant to DDT and permethrin. The difference between the least and the most tolerant to diflubenzuron was approximately two-fold. There was no correlation with resistance to the other insecticides. A DDT-resistant strain (T8) was selected 10 times (during 12 generations) with diflubenzuron. The LC₅₀ to diflubenzuron had increased 3.3 times by the S₈ generation but there was no further increase in later generations despite further selection. Associated with this increase, a marked decrease in resistance to DDT was observed but no change in permethrin tolerance. A genetically enriched strain (Hotchpotch) was synthesised from 35 strains of different geographic origin and crossed to the selected T8 strain before subsequent generations were selected five times with diflubenzuron. This procedure resulted in an 8 to 12-fold increase in the LC₅₀ value over that for unselected T8, accompanied by a decrease in the slope of the log dose against probit mortality line.     

苹果树腐烂病菌VmMFS1~VmMFS3基因的功能分析

为绿色持久防控苹果树腐烂病,该研究分析苹果树腐烂病菌Valsa mali的3个主要协同转运蛋白超家族（major facilitator superfamily,MFS）编码基因的氨基酸序列特征,利用实时荧光定量PCR （quantitative real-time PCR,RT-qPCR）技术分析这3个基因在苹果树腐烂病菌侵染阶段的表达水平,通过构建这3个基因的缺失突变体和回补菌株分析其在病原菌营养生长、致病力和非生物胁迫应答等方面的功能。结果表明,这3个基因的氨基酸序列均具有MFS保守结构域,将其命名为VmMFS1~VmMFS3; VmMFS1和VmMFS2的进化距离较近,均与VmMFS3的进化距离较远;在苹果树腐烂病菌侵染过程中VmMFS1~VmMFS3基因表达均显著上调;与野生型03-8菌株相比,VmMFS1~VmMFS3基因缺失突变体的菌落形态无明显差异,但生长速度下降; VmMFS1~VmMFS3基因缺失突变体的致病力均显著降低; VmMFS1~VmMFS3基因缺失突变体对H₂O₂胁迫的敏感性无明显变化,但对NaCl胁迫更敏感;基因回补后基因缺失突变体的表型缺陷能恢复到野生型菌株的水平。     

Strain Level Identification of Pseudomonads Using Denaturing Gradient Gel Electrophoresis of 16S-23S Spacer Region rDNA

Specific Pseudomonas strains were detected by PCR amplification of the 16S-23S rDNA spacer region followed by denaturing gradient gel electrophoresis (DGGE) to generate DNA banding profiles. In initial studies, two diverse sequence areas within the 16S-23S rDNA spacer region were located in five closely related Pseudomonas fluorescens and P. putida strains. DNA banding profiles of 16 different pseudomonads were generated using PCR primers flanking this region, followed by DGGE of the PCR products. Distinct banding profiles were observed for each strain, and specificity could be increased by designing additional primers within the spacer region. A specific primer (513-1) was used to selectively amplify and detect a plant-disease suppressive bacterium, P. fluorescens strain 513, in soil. Six field soils from different locations were used with the 513-1 primer to test the specificity of this technique. Five soils did not yield any gel bands, but one soil led to a faint 250-bp band, similar in size to that of P. fluorescens 513. Resolution of strain 513 from the indigenous strain in this soil was achieved by DGGE of the amplified DNA fragments. The results therefore demonstrate the utility of PCR-DGGE analysis for strain-specific identification of pseudomonads in environmental samples. Received 17 January 2000/ Accepted in revised form 10 May 2000     

葡萄生单轴霉重寄生菌F3的鉴定及防治效果测定

为明确从生长异常的葡萄霜霉层上分离获得的菌株F3对葡萄生单轴霉Plasmopara viticola的重寄生作用及对葡萄霜霉病的防治效果,通过光学和扫描电子显微镜观察菌株F3对葡萄生单轴霉的重寄生作用,并对菌株F3进行形态特征观察及28S rDNA序列分析,采用孢子囊萌发抑制和离体叶片点样法测定该菌株对葡萄霜霉病的防治效果。结果表明,菌株F3对葡萄生单轴霉表现出覆盖、缠绕的重寄生现象。结合形态特征与序列分析结果将该菌株最终鉴定为层生镰刀菌Fusarium proliferatum。菌株F3分生孢子悬浮液及其无菌发酵液对葡萄霜霉菌孢子囊萌发具有显著的抑制作用,抑制率分别为86.8%和83.1%;经菌株F3分生孢子悬浮液预防处理后的离体葡萄叶片发病率为10.9%,显著低于对照的发病率98.8%,防治效果达88.9%;菌株F3分生孢子悬浮液与葡萄霜霉病菌孢子囊混合后点样接种处理的叶片发病率为50.0%,也显著低于对照,防治效果为49.1%;而治疗处理的叶片发病率与对照间无显著差异。表明层生镰刀菌对葡萄生单轴霉具有重寄生作用,且对葡萄霜霉病具有较强的预防控制作用。     

Effects of Azoxystrobin on Mycotoxin Production in a Carbendazim-Resistant Strain ofFusarium sporotrichioides

Carbendazim-resistant (RS) and control (CS) strains ofFusarium sporotrichioides Sherb., previously developed in our laboratory, were exposed to graded concentrations of azoxystrob in in broth media under shake-culture conditions for 2, 3, 4 and 8 days. Azoxystrobin concentrations were 0, 1, 10 and 100 mg 1^-1 broth and cultures were incubated at a constant 25°C. Mycelial growth was significantly affected by strain (P<0.01), azoxystrobin concentration (P<0.001) and incubation time (P<0.001). Combined results for the four incubation times showed that CS yielded higher mycelial mass than RS (P<0.01) only in the absence of azoxystrobin. At fungicide additions of 1, 10 and 100 mg P^-1 mycelial growth was reduced (P<0.001) with minimal strain differences (P>0.05) at all three doses of azoxystrobin. Significant (P<0.05 or better) strain-fungicide interactions were recorded in trichothecene production following exposure to azoxystrobin. At 4 and 8 days of incubation, the 10 mg 1^-1 addition of azoxystrobin stimulated T-2 toxin synthesis (P<0.05) only in RS cultures. In contrast, T-2 toxin enhancement in CS cultures occurred only on day 8 but at a lower level of azoxystrobin (1 mg1^-1). Thus, the stimulation of T-2 toxin synthesis depended upon strain and azoxystrobin level. Production of diocetoxyscirpenol (DAS) was affected by a more complex set of interactions. Overall means showed that, in comparison with initial values (on day 2 or 3), DAS output maximized significantly(P<0.05) on day 4 in RS cultures and on day 8 in CS. Marked strain effects were observed on exposure to the 10 mg 1^-1 level of azoxystrobin. At this level, DAS production was enhanced in RS only after 4 (P<0.01 ) and 8 (P<0.05) days of incubation, while in contrast, CS reduced DAS production. As with T-2 toxin, DAS production in CS was stimulated (P<0.05 or better) only at low exposure levels of azoxystrobin. In the case of neosolaniol (NEO), however, the main effect of strain was significant (P<0.05), with CS producing consistently more of the mycotoxin than RS on day 4 of the experiment. At this point, the NEO:T-2 toxin ratio was also higher in CS (0.63) than in RS (0.12), a feature reported by us previously. In conclusion, the present investigation has shown for the first time that the development of resistance to one fungicide can affect trichothecene production inF. sporotrichioides on exposure to a second fungicide. These results have been incorporated into a new classification scheme for fungicide efficacy which is also presented in this paper. http://www.phytoparasitica.org posting Oct. 7,2001.     

Toxicity of spinosad to susceptible and resistant strains of house flies,Musca domestica

The toxicity of spinosad, a new insecticide derived from the bacterium Saccharopolyspora spinosa, was evaluated against susceptible and resistant strains of house fly (Musca domestica L.). Spinosad was highly toxic to house flies based on 72-h LD₅₀ values and the symptoms of poisoning were consistent with a neurotoxic mechanism of action. Spinosad was relatively slow acting, with the maximum toxicity noted at 72 h. Piperonyl butoxide and S,S,S,-tribu-tylphosphorotrithioate synergized the toxicity of spinosad by 3·0- and 1·8-fold, respectively, while diethyl maleate had no significant effect. These results suggest that there is a small degree of monooxygenase-mediated spinosad detoxification in house flies, while hydrolases may be only minimally important and glutathione transferases may have no role. There were no substantial levels of cross-resistance detected, except in the LPR strain where a low 4·3-fold cross-resistance was observed. The cyclodiene-resistant OCR strain was 2·7-fold more sensitive to spinosad than the susceptible strain (CS). These results suggest that cross-resistance may not be a limiting factor for the use of spinosad against house flies. © 1998 Society of Chemical Industry