Effect of Ifchit1 gene of Isaria fumosorosea on mortality,oviposition and oxidase activities of Bemisia tabaci

Isaria fumosorosea is one of important entomopathogenic fungi showed a good potential in controlling Bemisia tabaci. The effects of I. fumosorosea ⊿Ifchit1 mutant (Ifchit1 gene deletion mutant) on the mortality, oviposition, and host immunological response of B. tabaci, on Brassica campestris L. plant, were evaluated under laboratory conditions. The wild-type fungal strain infection significantly increased insect mortality and reduced the oviposition effeciency of B. tabaci, whereas the ⊿Ifchit1 mutant was much less effective, resulting in higher survival and ovipositing of B. tabaci. The activities of four insect enzymes were examined during a time course of fungal infection. Insect phenoloxidase, perioxidase, and catalase activities were decreased in whiteflies treated with the wild type and mutant I. fumosorosea strain at 12–36?h post treatment. However, these enzyme activities increased in fungal-treated whiteflies as compared to controls between 36 and 60?h post-infection, reaching peak values. Superoxide dismutase activity in fungal-treated whiteflies was higher than that in controls during the entire experimental time course examined. The overall enzyme activity profiles in ⊿Ifchit1 mutant-treated whiteflies were significantly different from wild-type strain treatments. Our results showed that loss of the Ifchit1 gene in I. fumosorosea affects whitefly mortality, ovipositioning and various antioxidant enzyme activities, providing new insights into the role of chitinases in I. fumosorosea-insect host–pathogen interactions.     

Joint action of the entomopathogenic fungus Isaria fumosorosea and four chemical insecticides against the whitefly Bemisia tabaci

The strain IfB01 of Isaria fumosorosea (Paecilomyces fumosoroseus) is a promising myco-insecticide candidate for control of Bemisia tabaci. Joint action of strain IfB01 and four chemical insecticides [spirotetramat (Spi), acetamiprid (Ace), imidacloprid (Imi) and thiamethoxam (Thi)] against the whitefly B. tabaci was evaluated using the cooperative virulence index (c.f.). Substantial synergistic actions were found in the mixtures of IfB01 conidia with Spi, Imi and Thi 2–4 days after treatment. The greatest synergism was recorded in the mixture of IfB01 (2.5 × 10⁶ conidia/L) and Imi (12.5 mg/L), which had a c.f. value of 184 at day 3 after treatment. Furthermore, the shorter LT₅₀ values recorded in mixtures indicated that effectiveness improved when the conidia were applied with the chemical insecticides. However, no substantial synergisms were recorded with Ace and from day 5 onwards.     

Agroecological basis for the design of biotechnological traps based on Isaria fumosorosea for the biological control of Bemisia tabaci in strawberry crops

Isaria fumosorosea is an entomopathogenic fungus that is used as a control alternative for nymphs and adults of Bemisia tabaci. Currently there are some commercial products, however, in greenhouse or field, these do not reach the levels of control as in the laboratory because the viability of the spores decreases as a result of the conditions of application of these products in situ. The objective of this work is to implement, through agroecological data, a system of biotechnological traps based on I. fumosorosea to increase the control efficiency mainly of adults of B. tabaci in strawberry greenhouses. One way to quantify the degree of infestation of a crop is the use of yellow traps, likewise to determine the spatial distribution of adults. The Taylor method [(1984). Assessing and interpreting the spatial distributions of insect populations. Annual Reviews of Entomology, 29, 321–357) was used in five different strawberry cultivation models, finding aggregate and regular distributions. Finally, once the crop model with the highest degree of infestation was selected, the designed traps were tested and mortalities were obtained between 50% and 90% in both the laboratory and the greenhouse. The biotechnological traps based on I. fumosorosea both in the laboratory and in the greenhouse had statistically the same effect as those used under the traditional method used in the field that is aspersion; therefore, this alternative method of application can be a tool important for the biological control of this pest.     

Host plants influence susceptibility of whitefly Bemisia tabaci (Hemiptera: Aleyrodidae) to the entomopathogenic fungus Isaria fumosorosea (Hypocreales: Cordycipitaceae)

We quantified the tritrophic effect of host plant on the susceptibility of the sweetpotato whitefly Bemisia tabaci (Genn.) to a fungal pathogen in the laboratory. Second-instar whiteflies reared on cucumber, eggplant, tomato and bean plants for six generations were exposed to conidial suspensions of Isaria fumosorosea isolate IF-1106. Our results did not detect differences in response (proportional survival or median lethal time, LT₅₀ days) among insect populations derived from different plants that were treated with 10⁷ conidia/ml. However, at concentrations ≤ 5×10⁶ conidia/ml, whiteflies reared on bean and tomato died significantly more quickly (i.e. LT₅₀ of 4–5 days) compared with cucumber and eggplant reared populations (5–7 days). Bean and tomato-reared populations were also more susceptible to mycosis (LC₅₀ ≈ 6 × 10⁵ conidia/ml) compared with those reared on cucumber (1.9 × 10⁶ conidia/ml) and eggplant (1.5 × 10⁶ conidia/ml). A separate study confirmed that this differential response of whitefly populations to I. fumosorosea was not explained by differences in deposition rate of conidia on leaf surfaces (i.e. a dosage effect). Our findings show that host plants affect the pathogenicity and virulence of a herbivore pathogen, but depend on the rate of exposure (inoculum) applied.     

An Isaria fumosorosea SCAR marker for evaluation of soil,insect, and airborne samples

Molecular detection markers are needed for ecological studies of entomopathogenic fungi. In this study, a sequence-characterized amplified region (SCAR) marker of Isaria fumosorosea was used to detect the fungus in soil, insects, and airborne samples. These were artificially added with different fungal conidial concentrations. Specificity and sensitivity were tested with semi-nested PCR using oligonucleotides E-AA/M-CTA₁₂₄ F and E-AA/M-CTA₁₂₄ R for the first amplification and E-AA/M-CTA₁₂₄ F and E-AA/M-CTA₁₀₃ R for the second amplification. Specificity assays showed a specific band of 103 bp for DNA samples from 10 I. fumosorosea strains used. Negative results were observed for DNA samples from other species of Isaria, including I. amoene-rosea, I. farinosa, and Paecilomyces carneus as well as with other entomopathogens such as Metarhizium acridum, M. anisopliae, M. majus, M. flavoviride Type E, and Lecanicillium lecanii. Sensitivity assays showed that the specific SCAR marker detected 10⁴ conidia from I. fumosorosea EH-511/3 that were artificially mixed with soil, from 1 to 10⁴ conidia artificially mixed with Galleria mellonella (Lepidoptera: Pyralidae), and from 10 to 10⁵ conidia in Melinex tape for airborne samples. The marker was also able to detect conidia in airborne samples from cotton wicks in a mini wind tunnel. These SCAR markers for I. fumosorosea had excellent specificity and sensitivity and are relevant tools for ecological studies of this fungus.     

Influence of the entomopathogenic fungus Isaria fumosorosea on Axinoscymnus cardilobus (Coleoptera: Coccinellidae) under laboratory conditions

Effects of entomopathogenic fungus Isaria fumosorosea on biological characteristics and life table parameters of Axinoscymnus cardilobus, a predator of whiteflies, were studied using five different conidial concentrations under laboratory conditions. Non-significant differences were observed among different fungal concentrations on the percent survival of all immature stages. The survival of A. cardilobus on different host plants did not differ significantly. The prey consumption of third instar A. cardilobus on Bemisia tabaci treated with different fungal concentrations differed significantly at different time intervals. The developmental periods for all immature stages (from eggs, first, second, third, fourth instar nymph and pupae up to emergence) among the treatments were not statistically significant when compared to that of control. In the present study, fecundity, longevity, egg viability and life table parameters of females were not statistically significant over the different concentrations. It can be concluded that control strategies tested are compatible to a greater extent and incorporation of these have promising prospects for control of whitefly.     

The entomopathogenic fungi Isaria farinosa (formerly Paecilomyces farinosus) and the Isaria fumosorosea species complex (formerly Paecilomyces fumosoroseus): biology,ecology and use in biological control

The entomopathogenic fungi, Isaria farinosa and Isaria fumosorosea were known as Paecilomyces farinosus and Paecilomyces fumosoroseus, for more than 30 years. Both fungi have a worldwide distribution and a relatively wide host range. While I. farinosa currently is of minor importance in research and as biocontrol agent, I. fumosorosea is regarded as a species complex, and various strains are successfully used for biocontrol of several pest insects, mainly whiteflies. During the past 40 years, numerous scientific papers on both fungal species covering various research areas were published. This is the first monographic review on the biology, ecology and use of I. farinosa and I. fumosorosea as biocontrol agents. In the following compilation, data on the identity, natural occurrence and geographical distribution, host range, production of metabolites, effect of abiotic and biotic factors (temperature, relative humidity, solar radiation, fungus, host plant) and on the use of these species in biocontrol experiments against pest insects, plant pathogens and nematodes in the laboratory and the field are summarised. Sections on their effects on non-target organisms and safety issues are also included. The aim of this review is to increase our knowledge on these entomopathogenic fungi.     

Characterisation of Isaria fumosorosea isolates and their virulence toward the Diamondback Moth,Plutella xylostella

Some morphological and physiological characteristics of an Isaria fumosorosea isolate with diminished virulence, IFCF01-D, and its parent isolate, IFCF01, were evaluated and laboratory bioassays were performed to assess their virulence against Plutella xylostella. The relationship among these traits and virulence against P. xylostella is discussed. There were no significant differences in conidial viability, spore production and the time required for 50% germination (GT₅₀). Spore viability after incubation for 24 h at 25°C was greater than 98% for both isolates tested. Spore production on potato dextrose agar after 14 days incubation at 25°C was 4.68 × 10⁸ and 4.59 × 10⁸ conidia/mL for IFCF01 and IFCF01-D, respectively. When exposed to high temperatures (40, 45, 50 or 55°C) through a water bath for 10 min, conidial germination ranged from 0.83% to 84.0% for IFCF01 and 0% to 86% for IFCF01-D. Germination rate showed a negative relationship with the exposure temperature for both isolates. The per cent germination of isolate IFCF01 24 h after ultraviolet (UV) radiation (18 W, 240–260 nm) varied from 0% to 92% and 0% to 81% for IFCF01-D. Germination rate and the exposure time exhibited a negative correlation for both isolates tested. Conidial surface hydrophobicity of IFCF01 (60%) was significantly higher than that of isolate IFCF01-D (53%). Subsequently, using the cicada exuviae as the substrate for enzymatic analysis, Pr1 and chitinase activity demonstrated the contrasting virulence traits: higher specific activities for the more virulent IFCF01 and lower enzymatic levels for isolate IFCF01-D.     

玫烟色棒束孢诱导的小菜蛾免疫响应表达谱分析

【目的】本研究旨在筛选小菜蛾Plutella xylostella应对玫烟色棒束孢Isaria fumosorosea侵染的免疫应答及其网络调控基因,以进一步探讨小菜蛾对玫烟色棒束孢的防御机制。【方法】采用第二代高通量测序技术RNA-seq,对处理后12 h的感染玫烟色棒束孢和健康(平行对照)的小菜蛾4龄幼虫转录组进行了测序,通过生物信息学分析对得到的差异基因进行了功能注释和分类,对差异基因参与的信号通路进行了分析。【结果】在感菌和健康小菜蛾幼虫转录组两个表达谱里,分别获得了12 346 987和12 315 210个clean reads,有60.93%和61.26%的reads分别能比对到参考基因库里,其中完美匹配(perfect match)的比例分别为32.15%和32.73%。共得到351个显著差异表达基因(differentially expressed unigenes,DEUs),上调表达基因有275个,下调表达基因有76个,与免疫防御反应潜在相关的基因有156个。GO(Gene Ontology)富集分析有102个DEUs分布到46个GO term里,KEGG(Kyoto Encyclopedia of Genes and Genomes)pathway富集分析结果显示,有132个DEUs显著富集在13个代谢通路(pathway)里。【结论】这些差异表达基因中,大部分编码潜在的与免疫识别及调控相关的基因,集中在能量代谢、疾病反应和防御反应等相关通路。研究结果为挖掘与玫烟色棒束孢侵染相关的小菜蛾免疫应答候选基因提供了重要数据库,也为阐明小菜蛾对玫烟色棒束孢的免疫机制奠定理论基础。     

Laboratory evaluation of a new strain CCM 8367 of Isaria fumosorosea (syn. Paecilomyces fumosoroseus) on Spodoptera littoralis (Boisd.)

Laboratory experiments were conducted to assess the efficiency of the new strain CCM 8367 of Isaria fumosorosea on different stages of the Egyptian cotton leafworm, Spodoptera littoralis (Boisd.), including soil treatments with pre-pupal and pupal stages. Treatments of larval instars showed a high susceptibility with 3rd, 5th and last instars to suspension of this fungus with concentration 5?×?10⁷ spores/ml. Larval mortality was over 90%. There were no significant differences (P?=?0.7929, F?=?0.2346) between instar treatments. The commercial fungus, PreFeRal® strain Apopka 97 of I. fumosorosea, which was used in comparison with this new strain caused mortality rates of between 63.33 and 90%. Statistically, differences between the effects of CCM 8367 strain and Apopka 97 were highly significant on the last instar (P?=?0.0064, F?=?6.479) and extremely significant on the 3rd instar (P?<?0.0001, F?=?13.29). No significant differences were recorded between the two strains on the 5th instar (P?=?0.0597, F?=?3.233). Fungal treatments with the late stage insects (end of the last instar or pupal stage) in soil yielded interesting results: the mortality rate on end of final instar larvae was 16.66% when treated with Apopka 97 and 83.33% when treated with CCM 8367. Soil containing pupae of S. littoralis that were inoculated with CCM8367 resulted in a high number of malformed adults, and the mortality rate was 64.52% (32.27% of malformed adults died and 32.25% of pupae fully infected by fungus). Only 3.23% of samples produced morphologically normal adults in this test. The results conclude that the strain of I. fumosorosea CCM 8367 has strong insecticidal effects on S. littoralis and has the potential to be implemented as a novel biocontrol agent.     

Host plant influences pathogenicity of Beauveria bassiana to Bemisia tabaci and its sporulation on cadavers

Laboratory bioassays were conducted to determine host plant effect on pathogenicity of the entomopathogenic fungus Beauveria bassiana (Balsamo) Vuill. (Ascomycota: Hypocreales) to the sweetpotato whitefly, Bemisia tabaci (Gennadius) (Homoptera: Aleyrodidae). Fourth instar B. tabaci reared on cucumber, tomato, melon, green pepper, potato, eggplant, marrow, cabbage, bean or cotton, were treated with 1 × 10⁷ conidia/ml B. bassiana EABb 93/14-Tp isolate. Mortality caused by B. bassiana significantly increased with time and it was significantly affected by the host plant on which the nymphs were reared. Mean mortality of nymphs 8 days after inoculation ranged between 52.3±7.3 for nymphs reared on cotton and 91.8±5.8 for nymphs reared on cucumber. Average survival times of nymphs treated with the fungal suspensions were also significantly influenced by the host plant, with a mean of 4.7±0.1 days for nymphs reared on cucumber, 6.6±0.2 days for cotton and 6.9±0.1 days for green pepper. The production of newly formed conidia was also affected by host plant and varied from 111000±8600 conidia/cadaver for nymphs reared on cotton to 597000±28000 conidia/cadaver for those reared on melon.     

烟粉虱在不同寄主作物上的种群动态及化学防治

20 0 1年在河北省廊坊市研究了烟粉虱Bemisiatabaci(Gennadius)在主要寄主作物棉花、大豆、花生和玉米上的种群动态和化学防治方法。研究结果表明 ,烟粉虱的种群数量在寄主的生育期内持续增长 ,在 8月 2 2日左右达到高峰 ,之后逐渐减少。烟粉虱虽然寄主范围广泛 ,但在每种寄主上的发生量并不一致 ,调查结果表明烟粉虱在 4种不同寄主上的种群数量比较为 :棉花 >大豆 >花生 ,玉米上没有发现烟粉虱的卵及若虫 ,只有极少量的成虫在植株上逗留。在药效试验中 1 0 %吡虫啉WP、2 5 %扑虱灵WP和 1 0 %高效氯氰菊酯LC在厂家推荐浓度下对烟粉虱成虫的防治效果分别为 2 0 0 7%～ 3 3 1 9% ,2 0 62 %～ 44 88%和 4 0 5 %～ 6 78% ;对若虫的防治效果分别为 5 1 91 %～ 64 46% ,5 4 3 0 %～ 63 99%和 1 6 73 %～ 2 6 97%。     

印楝素的杀虫活性及其对烟粉虱的驱避作用

印楝素(azadirachtin)被认为是最有发展前景的植物源杀虫剂。文章主要介绍印楝素对昆虫的接触驱避、产卵驱避、拒食、毒杀、降低生殖力、生长调节等作用及其作用机理。同时也论述印楝素对烟粉虱Bemisia tabaci(Gennadius)寄主选择、产卵、孵化、若虫发育的影响。     

Arachidonic acid production by Mortierella alpina grown on solid substrates

Mortierella alpina grown in solid state fermentations on cereal substrates gave up to 16% lipid in the final biomass. Arachidonic was at 50% of total fatty acids, with a yield of 36 mg/g of original substrate. Microbial lipid production was successfully scaled up to use 5-kg dry substrate batches.     

Effect of Bemisia tabaci honeydew and its carbohydrates on search time and parasitization of Encarsia bimaculata

Abstract:  The search time of Encarsia bimaculata Heraty et Polaszek was examined and recorded by using filter paper discs in Petri dish arenas impregnated with Bemisia tabaci (Gennadius) honeydew or its component carbohydrates. Our data show that E. bimaculata used honeydew as a contact kairomone to locate its host. Females responded quantitatively to the honeydew and the search time in the treated patches increased. This increase was concentration-dependent with regard to the amount of honeydew applied, but it levelled off 0.05 g/ml onwards. The response of E. bimaculata to honeydew decreased with increasing time after application and was significantly reduced 8 days after treatment. The parasitoid females were also arrested in patches treated with glucose, fructose, trehalulose and trehalose; trehalose elicited the highest response in the parasitoids. However, sucrose and low concentrations of melezitose did not elicit a significant effect. Glucose, sucrose, fructose, trehalose and trehalulose increased the longevity of the parasitoid females by a factor of 8.4, 8.1, 6.3, 6.1 and 4.2, respectively. Melezitose and 0.15 g/ml natural honeydew, however, had no effect on parasitoid lifespan. The effects of aqueous spray of honeydew sugars on egg to adult survivorship and parasitization of B. tabaci by E. bimaculata significantly differed when compared with controls only in cases where mixtures of glucose + fructose and trehalose and fructose + trehalose were applied.     

Host plant evaluation behaviour of Bemisia tabaci and its modification by external or internal uptake of imidacloprid

The behaviour of adult Bemisia tabaci Gennadius (Homoptera: Aleyrodidae) was observed on leaves of cotton, Gossypium hirsutum L., to study the process of host plant evaluation. Seven distinct behavioural elements were identified and recorded for a period of 20 min after initial contact with leaves. The behaviour of the whiteflies was observed on untreated leaves or on leaves treated with either foliar or systemically applied imidacloprid. Thus, contact with imidacloprid by whiteflies was either via the external cuticle or by ingestion, allowing determination of the effect of uptake route on host evaluation behaviour. Analysis of behavioural sequences and comparison of latencies, frequencies and durations of elements of behaviour between treatments revealed that host evaluation behaviour was similar on untreated and foliar treated leaves, but markedly different on those treated systemically. After their first probe, whiteflies on systemically treated leaves were much more active, spending significantly greater time walking and dabbing the labium tip onto the leaf surface. There was also a 50% reduction in the total time spent probing into systemically treated leaves, compared to the other treatments. This effect was the result of a shorter mean probe duration, and no effect of treatment was found on the frequency of probing. Pre-oviposition behaviour was significantly more common on the systemically treated leaves, although the number of eggs laid was similar on all treatments. These results also show that the host evaluation phase of adult female B. tabaci was dominated by probing. The behavioural sequence that leads to probing was affected significantly by imidacloprid when ingested, but not by direct contact with this compound on the leaf surface. The wider implications for the mechanisms of whitefly–host plant interactions are discussed.     

Diacetyl production and growth of Lactobacillus rhamnosus on multiple substrates

Lactobacillus rhamnosus is a heterolactic acid bacterium, which can be used to produce flavour compounds like diacetyl and acetoin. Various startegies have been applied to improve the growth rate and diacetyl yield. The use of multiple substrates affected growth as well as the yield of diacetyl. Growth on a medium containing glucose demonstrated a diauxic growth profile, with the second phase of growth being on the product, lactic acid. L. rhamnosus also grew on a medium containing citrate. Growth on medium containing glucose+citrate demonstrated simultaneous utilization of carbon sources. L. rhamnosus did not grow in a medium containing acetate and also did not co-metabolize it with glucose. Maximum specific growth rate ( _max) was found to increase in the case of simultaneous utilization of glucose+citrate (0.38 h^–1) as compared to glucose as the sole carbon source (0.28 h^–1). The yields of diacetyl were also found to increase for glucose + pyruvate and glucose + citrate (0.10 and 0.05 g g^–1 of glucose, respectively) as compared to glucose alone (0.01 g g^–1 of glucose). The productivity of diacetyl on medium containing glucose and citrate was double that of a medium containing only citrate, although the yields were comparable.     

A general and direct method for the solid phase synthesis of intramolecularly quenched fluorogenic protease substrates using a bifunctional coumarin derivative

A general method for the solid phase preparation offluorogenic peptide substrates or intramolecularly quenchedones (IQFS) is presented, using the highly fluorescentbifunctional coumarin derivative 7-amino-4-coumarinyl-acetic acid. The key feature of this method is theconjugation of H–Aca–OH through its carboxyl group on theresin, followed by the development of the peptide chainthrough its amino group, using standard Fmoc-derived solidphase peptide synthesis methodology. The 2,4-dinitrophenylgroup was used as quencher and introduced directly to theresin-bound peptides. The IQFSDnp–Lys–Pro–Ile–Cys–Phe–Ile–Lys–Leu–Aca–OH (2) andfour Dnp–X-Lys–Pro–Ile–Cys–Phe–Ile–Lys–Leu–Aca–OH (3–6), where X = Val, Lys, Ser and Glu at P₆ position,potential substrates for cathepsin D, were synthesized forproving the utility of the method. The compoundsH–Ile–Lys–Leu–Aca–OH (7),H–Lys–Pro–Ile–Cys–Phe–Ile–Lys–Leu–Aca–OH (8),H–Leu–Aca–OH (9), Dnp–Leu–Aca–OH (10) and Dnp-Leu-OH (11) were also synthesized for comparisonpurposes. The fluorescence properties of compounds 9and 10 were measured.     

Chitinase production by Beauveria felinaRD 101: optimization of parameters under solid substrate fermentation conditions

Major parameters affecting the production of chitinase by Beauveria felinaRD 101 under solid substrate fermentation conditions have been optimized. Wheat bran moistened with 100 MS-HCl medium adjusted to pH 5.0, inoculated with 1 × 10¹⁰ conidia g^–1 initial dry bran and incubated at 28 °C for 6 days produced maximum chitinase activity of 6.34 U g^–1 initial dry substrate.     

Production of pectinesterase and polygalacturonase by Aspergillus niger in submerged and solid state systems

Production of pectinesterase and polygalacturonase by Aspergillus niger was studied in submerged and solid-state fermentation systems. With pectin as a sole carbon source, pectinesterase and polygalacturonase production were four and six times higher respectively in a solid state system than in a submerged fermentation system and required a shorter time for enzyme production. The addition of glucose increased pectinesterase and polygalacturonase production in the solid state system but in submerged fermentation the production was markedly inhibited. A comparison of enzyme productivities showed that those determined for pectinesterase and polygalacturonase with pectin as a carbon source were three and five times higher by using the solid state rather than the submerged fermentation system. The productivities of the two enzymes were affected by glucose in both fermentation systems. The membranes of cells from the solid state fermentation showed increased levels of C18:1, C16:0 and C18:0 fatty acids. Differences in the regulation of enzyme synthesis by Aspergillus niger depended on the fermentation system, favoring the solid state over the submerged fermentation for pectinase production. Received 12 May 1997/ Accepted in revised form 19 September 1997