球孢白僵菌对斜纹夜蛾高毒菌株筛选与制剂的研发

斜纹夜蛾Spodoptera litura是一种危害极大的农业害虫,为了探索1条可持续、无污染的新型防治方法,我们通过标准喷塔法从12株球孢白僵菌Beauveria bassiana中筛选出1株对斜纹夜蛾2龄幼虫高毒力的菌株--球孢白僵菌Bb2860菌株.在喷雾浓度为(2592±275)孢子·mm-2的高浓度下,菌株Bb2860对斜纹夜蛾2龄幼虫在第7d的校正死亡率达82.43%±6.03%.不同制剂的室内生测试验表明,孢子水悬液和2种油剂(分别为不含稳定剂和含有稳定剂)在中浓度319～529孢子·mm-2的喷雾浓度下,7d后对斜纹夜蛾2龄幼虫的累计死亡率分别为54.8%、70.0%和76.7%;菌药协同作用为86.8%.另外,于2009年9月在安徽马鞍山进行自然条件下对花椰菜叶上斜纹夜蛾幼虫的田间小区药效试验.处理9d后,纯菌剂的相对防效为77.64%;低剂量和常规用量5%氟啶脲(抑太保)的相对防效分别为49.02%和89.70%;菌药混用的相对防效为91.13%.本研究显示出利用球孢白僵菌制剂及其与低剂量抑太保混用防治斜纹夜蛾的良好应用前景.     

Effect of Formulation and Application Method on the Efficacy of Aerial and Submerged Conidia of Metarhizium flavoviride for Locust and Grasshopper Control

A study was carried out to investigate the relative infectivity of aerial and submerged conidia of Metarhizium flavoviride to Schistocerca gregaria and Zonocerus variegatus. The effect of formulation and application method on initial infectivity and field persistence of these conidia was investigated. Strain IMI 330189 was highly virulent to S. gregaria but showed relatively low virulence to Z. variegatus. Direct contact with conidia from the initial spray application resulted in 100% mortality of S. gregaria for all formulation and application combinations. The mean survival time of infected locusts was significantly shorter for treatments using a knapsack sprayer containing submerged conidia in water plus 10 ml litre⁻¹ ‘Codacide’™ (seven days), than treatments with aerial conidia in oil using ULV techniques (8.9 days) or submerged conidia in modified (water plus adjuvants) ULV (MULV) (nine days) or in water-based (VLV) applications (9·3 days). Both aerial and submerged conidia persisted long enough in the environment to effect significant mortality via secondary pick-up of spray residue from vegeta-tion. Persistence was greatest in the ULV and MULV treatments, where the oil component of the formulations provided greater protection of the conidia from environmental stresses. The consequences of secondary pick-up of conidia from the different treatments on total mortality from a single application were examined using a simple host–pathogen model. This predicted that the ULV treatment would be much more effective than the other treatments under conditions where direct contact with the spray was limited. The results of these investigations are discussed in the context of development of optimum spray strategies for control of locusts and grasshoppers, and other pests, under different environmental conditions.     

球孢白僵菌对西花蓟马成虫的毒力及体表侵染的扫描电镜观察

本文在室内测定球孢白僵菌Beauveria bassiana CYT4菌株对西花蓟马Frankliniella occidentalis成虫的致死作用,并电镜观察分生孢子在西花蓟马成虫体表的侵染过程。结果表明：白僵菌的致死率随孢子浓度的增加而增加,1×109、1×108、1×107和1×106孢子.mL-1的孢悬液接种试虫第6d的致死率分别为83.57%、66.08%、49.78%和20.22%;0.04μL.mL-1和0.06μL.mL-1的多杀菌素与1×108孢子.mL-1白僵菌孢悬液混合后对试虫接种第5d的致死率分别为89.42%和95.76%,而单独施用菌液为58.19%,单独施用多杀菌素仅为45.55%和61.33%,表明二者混合能提高对西花蓟马成虫的致死率。扫描电镜观察发现,经白僵菌孢悬液处理的西花蓟马成虫虫体,在触角基节、复眼、足部褶皱区、刚毛窝、翅、产卵器和各体节连接处均有大量孢子附着和萌发,但腹部孢子量很少,尤其在节间膜处几乎无孢子;孢子在1h时即附着在体表,12h时已萌发,36h时有穿透体壁现象,48h后整个虫体被菌丝包被。     

Toxicity of the entomopathogenic fungus, Metarhizium anisopliae (Deuteromycotina: Hyphomycetes) to adult females of the blowfly Lucilia sericata (Diptera: Calliphoridae)

The fungus Metarhizium anisopliae (Metchnikoff) (Deuteromycotina: Hyphomycetes) is a natural component of soil flora world-wide and is a causal agent of the green muscardine diseases of insects. The use of this pathogen as a potential biocontrol agent against adult females of the blowfly, Lucilia sericata (Meigen) (Diptera: Calliphoridae) was considered. The blowfly L sericata is an economically important agent of cutaneous myiasis of sheep, particularly in northern Europe. Exposure of flies to suspensions of fungal spores by immersion, topical application or tarsal contact with treated surfaces all resulted in high levels of fatal infection. Spores suspended in silicone oil resulted in higher levels of infection (50-70%) than those formulated in a 0.3 glitre(-1) solution of the detergent Tween 80 (10-20%). Spore concentration had a significant effect on levels of infection of flies, with the highest levels of mortality (64%) resulting from a suspension of 1 x 10(7) conidia ml(-1). The duration of tarsal contact had a significant effect on levels of infection. Mean infection levels of 30% were observed following exposure of free-flying adults to a single spore-treated, 5 x 15 cm2 cloth surface suspended from the roof of a cage (30 cm cube). The implications of the results for the potential use of M anisopliae in the biocontrol of blowflies are discussed.     

防治假眼小绿叶蝉的虫生真菌高毒菌株的筛选及田间防治效果

茶小绿叶蝉Empoasca spp.是我国最重要的茶树害虫,几乎对所有化学农药都产生了很高的抗药性,亟需开发经济有效的生物农药。本研究通过对从茶小绿叶蝉上分离的1株爪哇棒束孢Isaria javanicus和4株自其它叶蝉上分离的球孢白僵菌Beauveria bassiana的胞外蛋白酶活性测定,初选出活性较高的爪哇棒束孢和两株球孢白僵菌;对假眼小绿叶蝉低龄若虫的生物测定结果显示,爪哇棒束孢毒力最强,分别比两株球孢白僵菌高2.33～7.26倍和6.08～27.26倍,在1×108孢子mL 1浓度条件下第13 d的致死率达100%,LT50为5.9 d。通过随机区组设计进行的田间防治试验结果表明,使用爪哇棒束孢菌株配制的IJOD-1悬乳剂以7.5×1013孢子hm 2的剂量常量喷施,对假眼小绿叶蝉虫口的防治效果在第11 d时为50.32%;第25 d时追加1次放菌,在第55 d时的防治效果达68.40%。     

Induction of phytoalexins in pigeonpea (Cajanus cajan) in response to inoculation with Fusarium udum and other treatments

Cajanol accumulation was monitored in four cultivars of pigeonpea, Cajanus cajan (L) Millsp, after inoculation with 2 x 10(6) conidia ml-1 of Fusarium udum Butler. Rapid accumulation was observed only in wilt-resistant cultivar ICP 9145. Another wilt-resistant cultivar, ICP 8863, had low levels of cajanol similar to those in wilt-susceptible cultivars ICP 2376 and Malawi local, indicating that rapid cajanol accumulation is not positively correlated with resistance to wilt in pigeonpea. A comparison of various inducing agents showed live conidia of the pathogen to cause more rapid accumulation than the other agents.     

Formulation of vegetable fat pellets with pheromone and Beauveria bassiana to control the larger grain borer,Prostephanus truncatus (Horn)

The use of hydrogenated rapeseed oil as a carrier for conidia of the entomopathogenic fungus Beauveria bassiana (Bals) Vuill was investigated as part of a research programme on the control of the larger grain borer, Prostephanus truncatus (Horn). Melting the oil, which is solid at temperatures below 32 °C, allows the incorporation of materials such as aggregation pheromones and conidia; sudden cooling produces solid fat pellets. In attraction tests conducted with pellets containing P truncatus aggregation pheromone, significantly higher numbers of beetles were attracted to pellets containing pheromone at a concentration of 4 ml litre⁻¹ compared to control pellets for at least four weeks when held in Petri dishes in the laboratory and for at least six weeks when pellets held in insect traps were exposed to outside conditions. The attraction was retained over a period of storage in glass bottles; pellets stored in the freezer or incubator (at −10 °C or 27 °C) attracted beetles according to the pheromone level for the duration of the work (14 and 13 months respectively). When pheromones and conidia were incorporated into the same pellets they could be stored in a freezer or refrigerator retaining over 80% viability after 51 weeks; those stored in an incubator at 27 °C showed significantly lower germination at 20.7–27.2% after the same time. There was an indication that the pheromone caused a slight reduction in the viability of conidia, although this may have been just a slight delay in the speed of germination. Rapid dose transfer from pellets with conidia with and without the addition of pheromone was demonstrated. Insects were exposed to pellets for 24 hours and 96–100% mortality was observed in treatments containing conidia within six days of exposure. © 1999 Society of Chemical Industry     

Distribution and efficacy of drip-applied metam-sodium against the survival of Rhizoctonia solani and yellow nutsedge in plastic-mulched sandy soil beds

The effects of metam-sodium application rate on soil residence time, spatial and temporal distributions of methyl isothiocyanate and pest control efficacy were studied in a Georgia sandy soil. Metam-sodium 420 g L(-1) SL was drip applied at rates of 147 and 295 L ha(-1) in plastic-mulched raised beds. Methyl isothiocyanate concentrations in soil air space were monitored from four preselected sites: 10 and 20 cm below the emitter, and 20 and 30 cm laterally away from the emitter at 3, 12, 24, 48, 72, 120 and 240 h after chemigation. A higher rate of metam-sodium application resulted in higher methyl isothiocyanate concentrations in the soil. Highest methyl isothiocyanate concentrations were found at 20 cm below the emitter, and lowest at 30 cm laterally away from the emitter. Methyl isothiocyanate concentrations decreased with time and distance from the emitter. Lower methyl isothiocyanate concentration x time product values at 20 and 30 cm away from the emitter resulted in lower mortalities of Rhizoctonia solani Kühn and yellow nutsedge (Cyperus esculentus L.). The results demonstrated that methyl isothiocyanate can be delivered at lethal doses with drip-applied water downward within the beds. Lateral diffusion of methyl isothiocyanate from the point of application did not reach biologically active concentrations to affect the survival of R. solani or yellow nutsedge. Further studies on the lateral distribution of methyl isothiocyanate in sandy soils are needed to circumvent this limitation.     

Transfer of indoxacarb among workers of Coptotermes formosanus (Isoptera: Rhinotermitidae): effects of dose, donor:recipient ratio and post-exposure time

Horizontal transfer of indoxacarb among workers of the Formosan subterranean termite, Coptotermes formosanus Shiraki, was examined under laboratory conditions. The effects of dose (0, 10, 20, 50, 100 or 200 ng AI per donor), donor:recipient ratio (1:1, 1:4 or 1:9) and post-exposure time (2, 4, 8, 16, 20 and 24 days) on lethal transfer of indoxacarb were investigated using a donor/recipient model in groups of 100 workers. Transfer of lethal doses from donors to recipients was evidenced by significant recipient mortality in 13 out of 15 treatments within 24 days post-exposure. Dosage significantly affected indoxacarb transfer. Higher doses resulted in greater recipient mortality than lower doses. The highest dose tested resulted in 100% death of recipients and donors within 20 days. A dose of 100 ng resulted in recipient mortalities ranging from 68 to 100%, whereas doses < or =50 ng killed <60% of recipients within 24 days. Donor:recipient ratio also had considerable effect on indoxacarb transfer. At 24 days post-exposure, greater recipient mortalities were observed at ratios > or =1:4 at doses > or =20 ng, but only at a ratio 1:1 at dose 10 ng. Recipient mortality increased significantly as post-exposure time increased. The higher the doses applied on donors, the shorter were the times required for the onset of recipient mortality to occur and for recipients to reach maximum mortality. Significantly greater recipient mortalities were not observed at doses 20 and 50 ng until more than 20 days post-exposure, indicating a delayed activity of indoxacarb. Possible transfer mechanisms are discussed.     

Enhanced efficiency of electrostatically charged insecticide aerosols

The bioefficacy of a standard domestic aerosol insecticide has been compared with that of a similar spray in which the droplets were electrostatically charged. The aerosol was charged without the need for an external power supply, by enhancing natural charge separation processes that occur during atomisation. The charge-to-mass ratio achieved was 1.1 x 10(-4) C kg-1, compared with 2.2 x 10(-5) C kg-1 for the standard aerosol. Efficiency was assessed for insects in free flight. A direct space-spray application method was used to study the bioefficacy of these aerosols on houseflies, Musca domestica, and an indirect space-spray method on houseflies and mosquitoes, Culex quinquefasciatus. Two levels of concentration of active ingredients were compared. At low concentrations of 1.57 g kg-1 of bioallethrin and 0.29 g kg-1 of bioresmethrin, the charged aerosol achieved a significant reduction in KDT50 (time at which 50% of the flies were knocked down), compared with the standard aerosol. In the direct spray application to houseflies, the KDT50 was reduced by 50%, while in the indirect spray the KDT50 was reduced by 40%. In the indirect spray of mosquitoes, the KDT50 was reduced by 22%. With higher active ingredient concentrations of 2.09 g kg-1 for bioallethrin and 0.39 g kg-1 for bioresmethrin, the charged aerosol also demonstrated faster knockdown than the standard, but the improvement was less marked. In the direct spray application to houseflies, the KDT50 was reduced by 21% compared with the standard aerosol, while in the indirect spray the KDT50 was reduced by 16%. In the indirect spray of mosquitoes, the KDT50 was reduced by 24%. The percentage mortality after 24 h was also increased with the charged aerosol. Charge carried by the droplets results in increased interception of the insecticide, as the droplets are attracted to the insects in flight and space charge effects within the aerosol could cause improved dispersion in the test chamber.     

莱氏野村菌对夜蛾科害虫防治潜力的室内评测

本文用喷雾法测定了5株莱氏野村菌Nomuraea rileyi对棉铃虫Helicoverpa armigera、甜菜夜蛾Spodoptera exigua和斜纹夜蛾Spodoptera litura 3种夜蛾科害虫的致病力。配制孢子悬浮液对3种夜蛾科害虫进行雾化接种,孢子附着量为188.9～4048.3孢子.mm 2。接种后幼虫置于（24±1）℃和12L∶12D条件下人工饲料饲养,每天观察直至化蛹。时间-剂量-死亡率模型分析显示：5株莱氏野村菌菌株接种量为188.9～4048.3孢子.mm 2时,斜纹夜蛾第10 d的侵染率为60%～95%;菌株Nr15对斜纹夜蛾显示出极高的毒力水平（LC50=266.7孢子.mm 2,4048.3孢子.mm 2浓度下的LT50=5.81 d）,第10 d的侵染率达95%。供试菌株对棉铃虫和甜菜夜蛾的致病力相对较弱,侵染率均不足50%。研究表明,莱氏野村菌菌株对夜蛾科害虫体现出明显的寄主专化性,且有明显的量效关系。     

Persistence and efficacy of two diatomaceous earth formulations and a mixture of diatomaceous earth with natural pyrethrum against Tribolium confusum Jacquelin du Val (Coleoptera: Tenebrionidae) on wheat and maize

Laboratory tests were conducted to assess the insecticidal and residual effects of three diatomaceous earth (DE) formulations, Insecto, PyriSec and SilicoSec, against Tribolium confusum Jacquelin du Val on wheat and maize. Quantities of wheat and maize were treated with the above formulations at 500, 1000 and 1500 mg kg(-1) and stored at 25 degrees C and 55% relative humidity (RH). Samples were taken on the day of storage and every 30 days until completion of a 360 day period of storage. Adults of T. confusum were exposed to these samples at 25 degrees C and 55% RH and the mortality was measured after 24 and 48 h and 7 and 14 days of exposure. Adult mortality was higher on wheat than on maize. At the beginning of the storage period, mortalities after 14 day exposure on maize treated with the highest rate were 60, 63 and 81% for Insecto, PyriSec and SilicoSec respectively, while on wheat the mortality was 100% for all DEs. On the same commodities 360 days after treatment the respective mortality figures for wheat were 99, 98 and 100%, while in the case of maize they did not exceed 7%. Generally, mortality at exposures < or =48 h decreased with increasing storage time. Furthermore, although mortality on wheat increased with dose, the increase in DE dose from 1000 to 1500 mg kg(-1) resulted in only a small increase in mortality. Thus a DE treatment of 1000 mg kg(-1) was shown to provide long-term protection of wheat against T. confusum, although higher DE application rates and exposure intervals are needed for a satisfactory level of protection of maize against this pest.     

Evaluation of the Francome MkII exhaust nozzle sprayer to apply oil-based formulations of Metarhizium flavoviride for locust control

Conidia of the fungus Metarhizium flavoviride were formulated in a paraffinic oil, ‘Shellsol’ T, and sprayed using the Francome MkII exhaust nozzle sprayer. Germination of the conidia collected from the spray was reduced by 30% as compared to unsprayed conidia. However, in bioassays, there was no detectable difference in virulence with conidia collected from the spray samples and unsprayed formulation. This indicated that, despite the recorded reduction in the concentration of active conidia, the efficacy of the formulation remained unchanged after passing through the exhaust nozzle sprayer. The droplet size spectra produced by the sprayer were investigated using the Malvern series 2600cc particle size analyser. The optimum droplets for locust control produced by this sprayer were generated by the number 1 nozzle (internal diameter 2·5 mm) with the number 1 restrictor ring (internal diameter 12.5 mm) sprayed at a pressure of 0·2 bar. The droplets thus produced had a volume median diameter of 58 μm when the nozzle protruded between 1 and 2 mm above the level of the restrictor ring. Of the droplets in the spray plume created by these conditions, 33% were between 50 and 100 μm, a range recommended as an achievable optimum for the ultra-low-volume application of Metarhizium flavoviride. The role of the exhaust nozzle sprayer as a tool for the application of M. flavoviride for locust control is discussed with reference to other vehicle-mounted ultra-low-volume sprayers. © 1997 SCI     

金龟子绿僵菌FM-03生物学特性及其对柑橘粉蚧的侵染

为明确金龟子绿僵菌FM-03菌株生物学特性及其对柑橘粉蚧的侵染效果,室内测定不同温度、pH、光周期、碳氮源对该菌株生长和产孢量的影响;观察柑橘粉蚧受该菌株侵染后的死亡趋势,构建该菌株对柑橘粉蚧的时间-剂量-死亡率（TDM）模型。结果表明,金龟子绿僵菌FM-03菌株培养最适的温度为25℃、pH 7、光周期12L：12D、碳源为乳糖、氮源为酵母膏,在此培养条件下菌落直径和产孢量分别可达6.17 cm和3.70×10⁹孢子/皿。柑橘粉蚧受菌株FM-03侵染后的死亡趋势随菌株浓度升高和侵染时间延长而上升,1×10⁸、1×10⁷和1×10⁶孢子/mL处理的升幅明显高于其他2个处理,其中在1×10⁸孢子/mL处理的累计校正死亡率最高可达82.02%;构建的TDM模型,通过Pearson卡方和Hosmer-Lemeshow检验,显示菌株FM-03对柑橘粉蚧的剂量效应随侵染时间的延长逐渐增强,时间效应随浓度的升高逐渐增强,侵染8 d后的LC₅₀和1.00×10⁸孢子/mL处理的LT₅₀分别可达6.03×10⁵孢子/mL和4.53 d。综上,金龟子绿僵菌FM-03菌株生长快、产孢量高,对柑橘粉蚧的致病力强,生防应用前景良好。     

Natural occurrence of and inoculation experiments withConidiobolus coronatus andConidiobolus sp. in glasshouse populations ofBemisia tabaci

Bemisia tabaci (Gennadius), the sweetpotato whitefly, has only one known entomophthoralean pathogen,Erynia radicans (Entomophthoraceae). Two new pathogens have been isolated recently from a glasshouse population of this pest:Conidiobolus coronatus and another, undescribed species ofConidiobolus (Entomophthorales: Ancylistaceae). Artificial inoculation experiments revealed that eggs ofB. tabaci are practically immune to infection by either species. Second-instar larvae are highly resistant, only high doses of conidia causing between <1% and 4.6% mortality. Adults were found to be much more susceptible. Doses of 60 conidia/mm² ofC. coronatus caused average mortalities ofca 95%. The maximum mortality of adults caused byConidiobolus sp. was much lower,ca 30%, at a dose of 210 conidia/mm². The incubation period (inoculation to death) for both species, under our experimental conditions, is very short: 18–24 h forC. coronatus and 30 h forConidiobolus sp. Both fungi produced loricoconidia (conidia metamorphosed into resting spores) on cotton leaves and other dry surfaces. This ability allowedConidiobolus sp. to remain viable for 17–21 days on cotton leaves, in glasshouse conditions and in the absence of hosts, whileC. coronatus persisted for 10–14 days.     

Effect of Germination Initiation on Competitive Capacity of Trichoderma atroviride P1 Conidia

ABSTRACT Trichoderma biocontrol isolates are most effective as highly concentrated inocula. Their antagonism to other fungi may be a result of pregermination respiration. In a nutrient-rich medium, almost all Trichoderma atroviride P1 (P1) conidia initiated germination processes and increased respiration, even in dense suspensions. When 1 x 10(7) P1 conidia/ml were coinoculated with 1 x 10(5) Botrytis cinerea conidia/ml, dissolved oxygen fell to <1% within 2 h and the pathogen failed to germinate. More dilute P1 suspensions consumed oxygen slowly enough to allow coinoculated B. cinerea to germinate. On nutrient-poor media, fewer P1 conidia initiated germination. Oxygen consumption by the inoculum and inhibition of B. cinerea were enhanced when P1 conidia were nutrient activated before inoculation. Pregermination respiration also affected competitive capacity of the antagonist on solid substrates, where respiratory CO(2) stimulated germination rate and initial colony growth. These parameters were directly correlated with inoculum concentration (R(2) >/= 0.97, P < 0.01). After initiating germination, Trichoderma conidia became more sensitive to desiccation and were killed by drying after only 2 h of incubation on a nutrient-rich substrate at 23 degrees C. These results indicate that nutrient-induced changes preceding germination in Trichoderma conidia can either enhance or decrease their biological control potential, depending on environmental conditions in the microhabitat.     

Strawberry Plant Extracts Stimulate Secondary Conidiation by Colletotrichum acutatum on Symptomless Leaves

ABSTRACT Conidial suspensions of Colletotrichum acutatum were prepared in 1:27, 1:45, and 1:81 (wt/vol) dilutions of an extract of strawberry (cv. Tristar) flowers or leaves in water. Strawberry leaves and plastic coverslips were sprayed with the conidial suspensions, incubated at 25 degrees C and continuous wetness for 48 h, and the number of conidia and appressoria were counted. In another experiment, leaves and coverslips were sprayed with a conidial suspension in water, incubated for 72 h to establish C. acutatum populations, and placed in a growth chamber under dry conditions for up to 6 weeks. At each sampling time, leaves and coverslips were sprayed with flower extracts, leaf extracts, or water, incubated for 48 h at 25 degrees C and continuous wetness, and the number of conidia and appressoria were counted. Flower extracts significantly (P 相似文献   

Influence of oil droplet size on the toxicity of bifenthrin on cotton to tobacco budworm (Heliothis virescens)

Sprays of bifenthrin and paraffinic oil mixtures were applied to cotton leaves (Gossypium hirsutum L.) to investigate droplet size effects on the mortality of Heliothis virescens (F.). An air-assisted nozzle applied mixtures at 9.4 litre ha ?1 spray rate with volume median diameters (VMD) of 96 and 337 μm, as verified with a Malvern laser diffraction analyzer. An hour after spraying, susceptible 3rd-instars walked for 2-min through wet droplets of spray mixtures applied at bifenthrin rates of 22, 45, 67, 90. and 112 g a.i. ha ?1. Mortality was assessed at 21, 26, 45, 50, 74, and 117h after exposure. Final mean mortality was affected (P =0.01) by bifenthrin rate and droplet size. Bifenthrin applied in 337 μm VMD droplets caused 75.8% mortality, whereas 96-μm VMD droplets resulted in a mortality of 67.9%. Also, the 337 μm VMD droplets killed larvae in 13% less time than the 96μm VMD droplets, based on analysis (P =0.01) using the mean elapsed time till death as the dependent factor.     

Effect of temperature,relative humidity,leaf wetness and leaf age on <Emphasis Type="Italic">Spilocaea oleagina</Emphasis> conidium germination on olive leaves

The effects of temperature, relative humidity (RH), leaf wetness and leaf age on conidium germination were investigated for Spilocaea oleagina, the causal organism of olive leaf spot. Detached leaves of five ages (2, 4, 6, 8 and 10 weeks after emergence), six different temperatures (5, 10, 15, 20, 25 and 30°C), eight wetness periods (0, 6, 9, 12, 18, 24, 36 and 48 h), and three RH levels (60, 80 and 100%) were tested. Results showed that percentage germination decreased linearly in proportion to leaf age (P < 0.001), being 58% at 2 weeks and 35% at 10 weeks. A polynomial equation with linear term of leaf age was developed to describe the effect of leaf age on conidium germination. Temperature significantly (P < 0.001) affected frequencies of conidium germination on wet leaves held at 100% RH, with the effective range being 5 to 25°C. The percent germination was 16.1, 23.9, 38.8, 47.8 and 35.5% germination at 5, 10, 15, 20 and 25°C, respectively, after 24 h. Polynomial models adequately described the frequencies of conidium germination at these conditions over the wetness periods. The rate of germ tube elongation followed a similar trend, except that the optimum was 15°C, with final mean lengths of 175, 228, 248, 215 and 135 μm at 5, 10, 15, 20 and 25°C, respectively after 168 h. Polynomial models satisfactorily described the relationships between temperature and germ tube elongation. Formation of appressoria, when found, occurred 6 h after the first signs of germination. The percentage of germlings with appressoria increased with increasing temperature to a maximum of 43% at 15°C, with no appressoria formed at 25°C after 48 h of incubation. Increasing wetness duration caused increasing numbers of conidia to germinate at all temperatures tested (5–25°C). The minimum leaf wetness periods required for germination at 5, 10, 15, 20 and 25°C were 24, 12, 9, 9 and 12 h, respectively. At 20°C, a shorter wetness period (6 h) was sufficient if germinating conidia were then placed in 100% RH, but not at 80 or 60%. However, no conidia germinated without free water even after 48 h of incubation at 20°C and 100% RH. The models developed in this study should be validated under field conditions. They could be developed into a forecasting component of an integrated system for the control of olive leaf spot.     

Evaluation of the time-concentration-mortality responses of Plutella xylostella larvae to the interaction of Beauveria bassiana with a nereistoxin analogue insecticide

Six bioassays were conducted to evaluate the interaction between Beauveria bassiana SG8702 and a nereistoxin analogue insecticide, diammonium S,S'-(2-dimethylaminotrimethylene)di(thiosulfate), which is highly compatible with the fungal biocontrol agent against diamondback moth Plutella xylostella (L.). Second-instar larvae were exposed to sprays of B. bassiana alone (assay 1) at concentrations of 21-38, 157-232 and 822-1133 conidia mm(-2) or together with the insecticide at the low application rates of 5, 10, 25, 50 and 100 microg AI ml(-1) (assays 2-6), and then maintained at 25 degrees C and 12:12 h light:dark photoperiod for daily monitoring of mortality for 8 days. Based on the modelling of the resultant time-concentration-mortality data sets, the fungal agent was highly virulent to P. xylostella with an LC50 decreasing from 269 conidia mm(-2) on day 4 to 107 on day 8. Lower lethal concentrations or shorter median lethal times resulted from fungal sprays including the tested chemical rates; the latter never caused higher mortalities than the fungal treatments alone. The fungal action over 3-7 days after spray was significantly enhanced by including in the fungal sprays the chemical at rates of > or =25 microg ml(-1) for 2.6- to 1756-fold reduction of LC50 values, > or =50 microg ml(-1) for 4- to 274-fold reduction of LC70 values and 100 microg ml(-1) for 9- to 33-fold reduction of LC90 values respectively. These rates were equivalent to 5-20% of the chemical rate labelled for field application. The fungal and chemical interaction outlined above highlights the feasibility of combined formulation or application of B. bassiana and the chemical insecticide for P. xylostella control.